You are on page 1of 133

BT0203 Genetics and Cytogenetics UNIT – I Terminologies Karyotype: A Karyotype is an array of chromosomes created by photographing the metaphase chromosomes

from one cell, cutting out the individual chromosomes from the photograph and lining them up in order from largest to smallest, pairing the appropriate homologous chromosomes. Chromosomes for karyotypes are often stained using special procedures which create banding patterns the chromosomes, thus making pairing easier. Diploid: A diploid nucleus contains two sets of chromosomes (two of each type of chromosome). Haploid: A haploid nucleus contains a single set of chromosomes. Polyploidy: Polyploids contain more than two sets of chromosomes in each nucleus. For example, bananas are triploid. Homologous chromosomes: Homologous chromosomes are the same size, the same shape, and have the same gene map. They are not necessarily genetically identical. Genome; A genome is an individual organism’s total array of genetic information. Gene pool: The gene pool is the total genetic diversity of a particular species. Gene: A gene is a segment of DNA which controls the production of a particular characteristic. More precisely, a gene is a recipe for the production of a specific kind of protein. Allele: Alleles are different forms of the same gene. For any gene, an individual may possess only two alleles, and a gamete may possess only one. However, the gene pool of a species may contain many alleles for any gene. Alleles are assigned symbols according Rex Arunraj Assistant Prof. Department of Genetic Engineering

BT0203 Genetics and Cytogenetics to specific rules of convention. All alleles of a particular gene should be given versions of the same symbol. Locus (plural Loci): A gene’s locus is its position on a chromosome. All genes have individual and unique locations characteristic of the species. What makes two organisms members of the same species is that they have the same assortment of genes, arranged according to the same gene map on their chromosomes. In other words, their various genes have the same loci. Multiple alleles: A gene has multiple alleles if there are more than two different alleles for that gene in the gene pool. For example, there are three different alleles for the A-BO blood type gene in human populations (LA, LB and l). Genotype: The genotype of an organism is the list of the symbols representing that organism’s specific genetic constitution—in other words, a list of all the alleles the individual carries for its genes. In actual usage, a stated genotype typically describes only one or two genes at a time. Phenotype: The phenotype is the actual physical expression of an organism’s traits. Much of the phenotype is the product of the genotype, but environmental influence can be very important as well. Geneticists discuss the heritability of traits. Heritability is the expression of the degree to which a particular trait is controlled by heredity. Homozygous; A homozygous individual has two identical alleles for the gene in question. For example, BB, bb, AA, PP. Heterozygous: A heterozygous individual has two different alleles for the gene in question. For example, Bb, Aa, Pp. Hemizygous: This term refers to the condition of a gene which is carried on an X or Y chromosome in a male. Since there can be only one copy of such a gene in the cell, the Rex Arunraj Assistant Prof. Department of Genetic Engineering

BT0203 Genetics and Cytogenetics terms homozygous and heterozygous are inappropriate. Essentially, hemizygous means that a gene is present in only one copy. Complete dominance: If two alleles display complete , it is not possible to tell the difference between the homozygous dominant individual and the heterozygous individual. The recessive allele is hidden by the presence of the dominant allele. Dominant: The dominant allele is the one which is displayed in the phenotype of the heterozygote. In assigning allelic symbols, the convention is to assign a capital letter to the dominant allele. Dominant traits can never skip generations in a pedigree, because they can never be present but hidden—they always show. Recessive: The recessive allele is the one which is hidden in the phenotype of the heterozygote. The recessive allele is generally assigned a lower case letter symbol. Recessive traits can skip generations in a pedigree. Incomplete dominance: If two alleles show incomplete dominance, the phenotype of the heterozygote is intermediate between the phenotypes of the two homozygotes. For example, RR produces red flowers, R’R’ produces white flowers, and RR’ produces pink flowers. Alleles showing incomplete dominance are typically assigned symbols which are variations of capital letters. Co-dominance: If two alleles show co-dominance, the phenotype of the heterozygote expresses both of the alleles completely. For example, in the A-B-O blood group, the LA and LB alleles show co-dominance. The heterozygote (Type AB) has all of the bloody type characteristics of Type A blood as well as those of Type B blood. Again, codominant alleles are generally assigned different versions of the same capital letter for symbols. Pseudodominance: Pseudodominance results when a particular genotype is lethal. For example, curly sings in Drosophila (fruit flies). The heterozygote has curly wings, the Rex Arunraj Assistant Prof. Department of Genetic Engineering

Independent Assortment: Genes which are not linked will behave independently of each other. By convention. it is often a mating between an offspring and an individual of the same genotype as one of the offspring’s parents. Department of Genetic Engineering . but upon closer examination is not. Back cross: Most literally. Eg: AaBb x AaBb Test cross: This is the mating between an individual of unknown genotype and a homozygous recessive individual (eg. Monohybrid cross: This is a mating between two individuals who are both heterozygous for the one gene which you are following.BT0203 Genetics and Cytogenetics homozygous straight has straight wings (wild type). Autosomal traits are traits carried on any of the chromosomes other than the sex chromosomes. B. Rex Arunraj Assistant Prof.x bb) for the purpose of exposing hidden recessive alleles in the unknown parent. and will assort according to simple rules of probability. this is the mating between an offspring and one of its parents. Eg: Aa x Aa Dihybrid cross: This is a mating between two individuals. Linkage: Genes which are carried on the same chromosome are considered to be linked. Superficially. both of whom are heterozygous for the two genes you are following. the effect looks like complete dominance (the curly allele appears to be dominant). the homozygous lethal allele is given a capital letter symbol and the pseudo-recessive allele is given a lower case letter symbol. In practice. When crosses using two linked genes are made. and the homozygous curly is lethal (the eggs never hatch). and results do not behave according to simple rules of statistics. the two genes do not behave independently.

since an XY zygote always inherits its X from its female parent and its Y from the male parent. Eg. In mammals. males inherit all of their X-linked traits from their mothers. Sex influenced traits: Sex influenced traits are autosomal traits whose expression is affected by gender. Since this hormone is found in much higher levels in males than in females. but she can’t possibly express the trait. and all of their sons.BT0203 Genetics and Cytogenetics Sex linked traits: This is a special type of linkage. Since females have two copies of the X chromosome and males have only one. Y-lined traits found in a father must appear in all of his sons. pattern baldness in humans is sex influenced. A female can be genetically cryptorchid. males inherit all Y-linked traits from their fathers. maleness is carried on the Y chromosome. Rex Arunraj Assistant Prof. Y-linked (holandric) genes are carried on the Y chromosome. Sex limited traits are autosomal traits whose expression is possible only in one of the genders. These traits generally affect the primary and secondary sexual characteristics. by an autosomal gene. cryptorchidism is a condition in males in which one or both testes fail to descend into the scrotum late in gestation. Department of Genetic Engineering . it would be the Z or W chromosome). Sex linked genes are carried either on the X or the Y chromosome (in mammals or fruit flies—in birds. etc. X-linked genes are carried on the X chromosome. the alleles are influenced by the presence of certain hormones which either increase or decrease the effects of the alleles. Since Y chromosomes are inherited exclusively through the male line. Also. rare recessive characteristics which are Xlinked will occur more often in males than in females. The effectiveness of the bald allele is greatly increased in the presence of high levels of the hormone testosterone. the bald allele and the non-bald allele. The gene for this trait has two alleles. This characteristic is genetically controlled. Usually. the bald allele is dominant in males and recessive in females. Eg.

the white spotting gene in gerbils apparently also influences red blood cell count. Rex Arunraj Assistant Prof. Department of Genetic Engineering . Another simple example is that genes which influence characteristics of the fingers will also influence characteristics of the toes.BT0203 Genetics and Cytogenetics Pliotropy refers to genes with multiple effects. For example.

Mendel was not the first to conduct hybridization experiments.. Department of Genetic Engineering . 8. but just the extension of the experiments conducted by earlier worker. Dawis. of experiments with pea plants. The pea plant has contrasting characters. There was another scientist Kolreuter – A German Botanist performed experiments is tobacco similar observations were made by a group of workers Garther. Short growth period & growth cycle. He studied the inheritance of only one character at a time. Cross pollination was not very difficult. 6. 2. With this experiments he was able to explain the inheritance of characters. 7. His paper “experiments in plant hybridization were published is 1866 to 1867 in the proceeding of Natural History Society of Brunn. Pure breeding varieties were available of easy to cultivate. Reason’s for Mendel’s Success 1. which helped him to derive numerical ratio’s of significance. but they could not figure out their results numerically as Mendel.BT0203 Genetics and Cytogenetics Mendelian Genetics Gregor Johann Mendel is called the father of genetics. Naudin etc. He was fond of gardening and interested in plant hybridization and he performed a No. 3. He maintained statistical records of the results. Like Knight & Goss. 4. Carl Correns. His work was recognized only in 1900 by Hugo de Veries – a Dutch biologist. The genes for the seven pairs of characters are located on seven separate homologies pairs of chromosomes 5. The flower of pea plants are normally self fertilized. His work remained unnoticed for 33 years. Rex Arunraj Assistant Prof. a German botanist & Erich Von Tschermak an Austrian Botanist.

Reciprocal Cross T. F4 etc. The progeny of F1 plants obtained due to self pollinations is called second fillial generations or F2. The population obtained as a result of crossing plants exhibiting contrasting characters is called the first fillal generation or F1 progeny. Rex Arunraj Assistant Prof.tall female plant X t – dwarf male plant. Reciprocal crosses were also made.tall male plant X t –dwarf female plant. 5. and then they are self pollinated. 3. No. Character Alternatives Dominant 1. T. Length of the stem Position of the flower Colour of the pod Shape of the pod Shape of Seed Colour of the Seed coat Colour of the cotyledon Tall Axial Green Inflated Round Coloured Yellow Recessive Dwarf Terminal Yellow Constricted Wrinkled White Green Plants with one alternative trait were used as female and those with the other alternative as male. Similarly we can have F3. 7. Department of Genetic Engineering . 2. the anthers have to be removed and are called as emasculation. This stigma is protected against any foreign pollen grain. Character selected by Mendel Mendel selected seven characters with contrasting alternatives.BT0203 Genetics and Cytogenetics CROSSING TECHNIQUE Garden pea is self fertilizing. 6. 4.

showed only one of the traits & never the other. This feature was expressed as dominant of one trait over another. individuals from crosses. (i) For any character the F1. between two different varieties having alternative characters. When the F1. all plants is the F1. The tall and dwarf plants were obtained in the ratio of 3:1 similar patterns were obtained for other six pairs of character also. generation were tall. To summarize the pattern of inheritance in all the seven cases. both tall & dwarf plants were obtained in the F2 generation.BT0203 Genetics and Cytogenetics Results of Mendel’s experiments When tall plants were crossed with dwarf plants. The plants used are the initial crosses are referred to as P1 & P2 (or) parents. Department of Genetic Engineering . Rex Arunraj Assistant Prof. population was called recessive. the reciprocal crosses gave the same results. plants are self fertilized. The trait which appeared is the F1 generation was called dominant and the other which did not appear is the F1. (ii) It did not matter which parent variety provided the pollen which provided the eggs the results were always the same in other words. The determining agent responsible for each trait was called a factor.

In the F2 generation two types of plants were found.BT0203 Genetics and Cytogenetics Monohybrid Cross Single character each controlled by a single pair of genes or alleles were considered such crosses are known as monohybrid crosses the F2 ratio 3 :1 is known as monohybrid ratio. He could raise 1064 plants in F2 generation 787 plants were tall and 277 plants were always 75% tall plants 25% dwarf Plants. A pure breeding tall & dwarf plants were treated as pants & were crossed. The seeds were collected to the next generation was raised. Monohybrid Experiment The crossing of two plants differing in one character is called monohybrid experiment. F2. generation all the F1 plants were tall. They were tall and dwarf. tt t Department of Genetic Engineering . Parents P Gametes F1 F1 Selfing Gametes F2 Phenotype Tall TT T Tt Tt T TT 3:1 Gametes T t T TT Tall Tt Tall Rex Arunraj Assistant Prof. These seeds were sown and a group of plants were raised these plants constituted the first filial generations or F1. plants crossed. t Tt Tall Tt Dwarf t X X Tt T t Tt Tt tt X Dwarf. The F1. A pure breeding plant is one that which retains a particular character for any number of generations.

Hence each gamete will contain only allele. the pure dwarf plant two receive alleles tt. During gamete formations. recessive allele t.T. the domination allele T masks the effect of the Rex Arunraj Assistant Prof. Dominant . The resulting F1. Department of Genetic Engineering . 50% t. the alleles separate to enter two gametes.allele fuses with t.allele. The gametes of the F1 generation are 50% T. plant is -Tt.BT0203 Genetics and Cytogenetics Genotype 1:2:1 The pure tall plant has two dominant alleles for height TT. The gametes produced by a homozygous tall plant contain only one type of allele. It this plant. When a tall plant and dwarf plant are crossed the gametes containing T. recessive – t.

BT0203 Genetics and Cytogenetics Dihybrid Experiment The crossing of two plants differing in two characters is called dihybrid experiment. Two characters are considered at a time (Colour & Shape). Colour of the cotyledon – (Yellow & Green), seed shape – (Round & Wrinkled). Mendel selected a pure breeding yellow, round (Dominant) and a Pure breeding green, wrinkled seed producing plant ( Recessive ).The F1 generation plants produced only yellow round seeds. The F1 plants were self fertilized. In F2 generation four kinds of plants were produced. They are plants producing yellow, round seeds Plants producing yellow, wrinkled seeds Plants producing green, round seeds Plants producing green, wrinkled seeds Yellow (Y) is dominant over green (y); round seed shape (R) is dominant over wrinkled (r). The dominant parent produce only one type of gamete and each gamete are carrying one allele for colour (Y) & another allele for seed shape (R). The F1 plants Yellow and Round YyRr When F1 hybrid is selfed Gametes YR YR YYRR yellow round Yr YYRr yellow round yR YyRR Yr YYRr yellow round Yyrr yellow wrinkled YyRr yR YyRR yellow round YyRr yellow round yyRR yr YyRr yellow round Yyrr yellow wrinkled yyRr

Rex Arunraj Assistant Prof.

Department of Genetic Engineering

BT0203 Genetics and Cytogenetics yellow round Yyrr Yellow wrinkled Dihybrid ratio: 9:3:3:1 Mendel’s Laws Based on Mendel’s experiments results certain principles are framed. These are called Mendel’s laws. 1. 2. 3. Law of dominance Law of segregation or law of purity of gametes. Law of independent assortment yellow round Yyrr Yellow wrinkled green round yyRr green round green round Yyrr green wrinkled


Law dominance Each organism is made of a bundle of character; each character is controlled by factors or alleles or genes. Mendel’s law of dominance states that one factor in a pair may mask or prevent the expression of the other. He called the variety that appeared in the F1 generation of a monohybrid cross as dominant variety and that which did not appear in F1 generation to be recessive. A recessive factor freely expresses itself with the absence of the dominant allele. Law of segregation Each character is controlled by a pair of alleles. The two alleles of a particular character remain uncontaminated when they are inside the organism during gamete formation the paired alleles segregated & enter different gametes. During gamete formation the alleles of particular character separate and enter different gametes. segregation. formulated based on monohybrid experiments. Mendel made two innovations to the science of genetics: Rex Arunraj Assistant Prof. Department of Genetic Engineering This is the law of These laws were This law is also called law of purity of gametes.

BT0203 Genetics and Cytogenetics • • developed pure lines counted his results and kept statistical notes

Pure Line - a population that breeds true for a particular trait [this was an important innovation because any non-pure (segregating) generation would and did confuse the results of genetic experiments] Results from Mendel's Experiments Parental Cross Round x Wrinkled Seed Yellow x Green Seeds Red x White Flowers Tall x Dwarf Plants F1 Phenotype Round Yellow Red Tall F2 Phenotypic Ratio 5474 Round:1850 Wrinkled 6022 Yellow:2001 Green 705 Red:224 White l787 Tall:227 Dwarf F2 Ratio 2.96:1 3.01:1 3.15:1 2.84:1

Phenotype - literally means "the form that is shown"; it is the outward, physical appearance of a particular trait Mendel's pea plants exhibited the following phenotypes: - round or wrinkled seed phenotype - yellow or green seed phenotype - red or white flower phenotype - tall or dwarf plant phenotype Dominant - the allele that expresses itself at the expense of an alternate allele; the phenotype that is expressed in the F1 generation from the cross of two pure lines Recessive - an allele whose expression is suppressed in the presence of a dominant allele; the phenotype that disappears in the F1 generation from the cross of two pure lines and reappears in the F2 generation Rex Arunraj Assistant Prof. Department of Genetic Engineering

The F1 from a cross of two pure lines contains one allele for the dominant phenotype and one for the recessive phenotype. Each parent has a allele pair in each cell for each trait studied. These two alleles comprise the allele pair. Phenotypes F2 Tall Plants Rex Arunraj Assistant Prof. And indeed. From these results we can now confirm the genotype of the F2 individuals. If his law was correct he could predict what the results would be. the results occurred has he expected. To test this hypothesis. Mendel could form a hypothesis about segregation. Mendel's First Law . Genotypes Genetic Description 1/3 DD Pure line homozygote dominant Department of Genetic Engineering . These determinants are called genes. Mendel selfed the F2 plants. Gametes unite at random and irrespective of the other allele pairs involved.BT0203 Genetics and Cytogenetics Mendel's Conclusions The hereditary determinants are of a particulate nature. One member of the allele pair segregates into a gamete. during gamete formation each member of the allelic pair separates from the other member to form the genetic constitution of the gamete Confirmation of Mendel's First Law Hypothesis With these observations. thus each gamete only carries one member of the allele pair.the law of segregation.

the offspring of two parents that are homozygous for alternate alleles of an allele pair Rex Arunraj Assistant Prof. the first cross is between two pure line parents to produce an F1 heterozygote. Backcross: Dd x dd Backcross . Department of Genetic Engineering . Monohybrid cross . So although the phenotypic ratio is 3:1 the genotypic ratio is 1:2:1 Mendel performed one other cross to confirm the hypothesis of segregation --. Mendel crossed it to a pure line. all the discussion has concentrated on monohybrid crosses. used to determine if the individual is homozygous dominant or heterozygous So far. though a backcross is a cross to a fully recessive parent Testcross .the backcross. At this point instead of selfing the F1.the cross of any individual to a homozygous recessive parent. most often.a cross between parents that differ at a single allele pair (usually AA x aa) Monohybrid . homozygote dwarf plant. for pea plant height the cross would be Dd x DD or Dd x dd.BT0203 Genetics and Cytogenetics 2/3 Dd F2 Dwarf Plants all dd Heterozygotes Pure line homozygote recessive Thus the F2 is genotypically 1/4 Dd : 1/2 Dd : 1/4 dd This data was also available from the Punnett Square using the gametes from the F1 individual. Remember.the cross of an F1 hybrid to one of the homozygous parents.

the ability of one allele to express its phenotype at the expense of an alternate allele. According to this law the alleles for each pair of character separate independently from those of other character during gametes formation.BT0203 Genetics and Cytogenetics Monohybrids are good for describing the relationship between alleles. generally the dominant allele will make a gene product that the recessive can not. It is the phenotype of the heterozygote which permits us to determine the relationship of the alleles. First. Parental Cross: Yellow. the law of independent assortment. These experiments formed the basis of his discovery of his second law. Dihybrid cross . the major form of interaction between alleles. Dominance . Mendel also performed crosses in which he followed the segregation of two alleles. Now. a few terms are presented. Department of Genetic Engineering . When an allele is homozygous it will show its individual heterozygous for two pairs of alleles (AaBb) Again a dihybrid cross is not a cross between two dihybrids. To this point we have followed the expression of only one allele. round Rex Arunraj Assistant Prof. During gametes formations the allele Y may combine with the dominant allele R or recessive allele -r of the other character. let's look at a dihybrid cross that Mendel performed. Wrinkled Seed F1 Generation: All yellow.a cross between two parents that differ by two pairs of alleles (AABB x aabb) Dihybrid. therefore the dominant allele will express itself whenever it is present Law of independent assortment This law is based on dihybrid experiments. Round Seed x Green.

Now set up the Punnett Square for the F2 cross. Gw GGWw GGww (Yellow. Green = g Seed Shape: Round = W. let's diagram the cross using specific allele symbols. (Yellow. (Yellow. Wrinkled. Wrinkled = w The dominance relationship between alleles for each trait was already known to Mendel when he made this cross. The purpose of the dihybrid cross was to determine if any relationship existed between different allelic pairs. (Yellow. Wrinkled At this point. Round.BT0203 Genetics and Cytogenetics F2 Generation: 9 Yellow. (Yellow. (Yellow. Choose Symbol Seed Color: Yellow = G. 3 Yellow. Female Gametes GW Gw gW gw GGWW GGWw round) Male round) GgWW GgWw round) GgWw round) Ggww GW (Yellow. 1 Green. Department of Genetic Engineering . (Yellow. Rex Arunraj Assistant Prof. Round. 3 Green. Let's now look at the cross using our allele symbols.

F1 dihybrid x recessive parent. during gamete formation the segregation of the alleles of one allelic pair is independent of the segregation of the alleles of another allelic pair As with the monohybrid crosses. Wrinkled Seed 3 Green. round seeded F1. Let's use the example of the yellow. (Green. wrinkled) round) The phenotypes and general genotypes from this cross can be represented in the following manner: Phenotype 9 Yellow. (Green. Round Seed 3 Yellow. Department of Genetic Engineering .the law of independent assortment. Mendel confirmed the results of his second law by performing a backcross .BT0203 Genetics and Cytogenetics round) Gametes wrinkled) round) ggWW round) ggWw wrinkled) ggWw (Green. wrinkled) GgWW GgWw round) GgWw gw round) round) Ggww gW (Yellow. Wrinkled Seed General Genotype G_W_ G_ww ggW_ ggww The results of this experiment led Mendel to formulate his second law. (Yellow.ROUND) ggww (Green. (Yellow. Mendel's Second Law . Rex Arunraj Assistant Prof. (Yellow. Round Seed 1 Green.

Department of Genetic Engineering . Wrinkled Seed 1 Green. Round Seed 1 Yellow. a statistical test called the goodness-of-fit chi-square test is used. This test provides information about how well observed values fit expected values. round) (Green. we need some means of evaluating how likely it is that chance is responsible for the deviation between the observed and the expected numbers. Before we learn how to calculate the chi square. or whether we have chosen the correct genetic explanation Rex Arunraj Assistant Prof. Wrinkled Seed Testing for Independent Assortment The Goodness-of-Fit Chi-Square Test Clearly. To evaluate the role of chance in producing deviations between observed and expected values. wrinkled) The phenotypic ratio of the test cross is: • • • • 1 Yellow. whether the results are correct. The chi-square test cannot tell us whether a genetic cross has been correctly carried out.BT0203 Genetics and Cytogenetics Punnett Square for the Backcross Female Gametes GW Male Gametes Gw gW gw Ggww ggWw ggww gw GgWw (Yellow. it is important to understand what this test does and does not indicate about a genetic cross. wrinkled) (Green. Round Seed 1 Green. round) (Yellow.

not to proportions or percentages. In other words. Most scientists use the . Department of Genetic Engineering .BT0203 Genetics and Cytogenetics for the results. where n is the number of different expected phenotypes. The chi-square test must always be applied to numbers of progeny. We are ready to obtain the probability from a chi-square table (Table). When the probability calculated from the chi-square test is high. we assume that some factor other than chance—some significant factor—produced the deviation. The degrees of freedom are given in the left hand column of the table and the probabilities are given at the top. The chi-square value is calculated by using the following formula: χ2 = Σ (Oserved – Expected) 2 / Expected where Σ means the sum of all the squared differences between observed and expected divided by the expected values. What it does indicate is the probability that the difference between the observed and the expected values is due to chance.05 probability Rex Arunraj Assistant Prof. which is the probability that the deviation between the observed and the expected results could be due to chance. When the probability is low. The theoretical chi-square values increase from left to right and the probabilities decrease from left to right. it indicates the likelihood that chance alone could produce the deviation between the expected and the observed values. This step requires us to compare the calculated chi-square value with theoretical values that have the same degrees of freedom in a chi-square table. we first determine the expected results. Find where calculated chi-square value lies among the theoretical values in this row. The next step is to determine the probability associated with this calculated chi-square value. The degrees of freedom represent the number of ways in which the observed classes are free to vary. we assume that chance alone produced the difference. within the body of the table are chi-square values associated with these probabilities. find the row for the appropriate degrees of freedom. To use the goodness-of-fit chi-square test. For a goodness-of-fit chi-square test. the degrees of freedom are equal to n-1. First.

straight wings Total = Rex Arunraj Assistant Prof. straight wings 77 yy cvcv yellow body.BT0203 Genetics and Cytogenetics level as their cutoff value: if the probability of chance being responsible for the deviation is greater than or equal to . curved wings 32 yy cv+cv yellow body. Suppose we did a testcross for two pairs of genes. For example. To illustrate this analysis. Is this outcome a 1:1:1:1 ratio? Not exactly. curved wings 28 y+y cvcv brown body. and so the number of nonrecombinants is only slightly greater than the number of recombinants. in which yellow body (y) is recessive to brown body (y+) and curved wings (cv) are recessive to straight wings (cv+). 42 Aabb. How do we distinguish between the roles of chance and of linkage in producing deviations from the results expected with independent assortment? Testing for independent assortment between two linked genes requires the calculation of a series of three chi-square tests. scientists assume that chance is not responsible and a significant difference exists. with considerable crossing over taking place between them. 200 total progeny Department of Genetic Engineering . When the probability is less than .05. and observed the following numbers of progeny: 54 AaBb. Alternatively. A testcross (y+y cv+cv X yy cvcv) produced the following progeny: 63 y+y cv+cv brown body. the genes might be linked. and 48 aaBb. 56 aabb. such as AaBb X aabb. The expression significant difference means that some factor other than chance is responsible for the observed values being different from the expected values.05. we will examine the data from a cross between German cockroaches. Perhaps these genes are assorting independently and chance produced the slight deviations between the observed numbers and the expected 1:1:1:1 ratio. they accept that chance may be responsible for the deviation between the observed and the expected values. but its pretty close.

where n equals the number of expected classes.1 = 1. so the degree of freedom is 2 . We observe 63 + 28 = 91 brown progeny and 77 + 32 = 109 yellow progeny. we conclude that there is no significant difference between the 1:1 ratio that we expect in the progeny of the testcross and the ratio that we observed. Looking up our calculated chi-square value in Table. we obtain: The degrees of freedom associated with the chi-square test are n. so we expect 100 of each. Because the probability is above . Rex Arunraj Assistant Prof.30 and . Here. At the first locus (for body color). there are two expected phenotypes. the cross between heterozygote and homozygote (y+y X yy) is expected to produce half y+y brown and half yy yellow progeny.20.BT0203 Genetics and Cytogenetics Testing ratios at each locus To determine if the genes for body color and wing shape are assorting independently. we find that the probability associated with this chi-square value is between . Department of Genetic Engineering . Applying the chi-square test to these observed and expected numbers.1. we must examine each locus separately and determine whether the observed numbers differ from the expected (we will consider why this step is necessary at the end).05 (our critical probability for rejecting the hypothesis that chance produces the difference between observed and expected values).

so the calculated chi-square value is: The degree of freedom associated with this chi-square value also is 2 . At this locus.BT0203 Genetics and Cytogenetics We next compare the observed and expected ratios for the second locus.1 = 1. Department of Genetic Engineering .5 and . We actually observe 63 + 32 = 95 straightwinged progeny and 77 + 28 = 105 curved-wing progeny.3. Testing ratios for independent assortment Rex Arunraj Assistant Prof.We again assume that there is no significant difference between what we observed and what we expected at this locus in the testcross. a heterozygote and homozygote also were crossed (cv+ cv X cvcv) and are expected to produce half cv+cv straight-winged progeny and half cvcv curved-wing progeny. which determines the type of wing. and the associated probability is between .

Our conclusion. If the genes are assorting independently. followed by a test for independent assortment between alleles at the different loci.1 = 3 and the associated probability is considerably less than . testing for linkage between two genes requires a series of chi-square tests: a chi-square test for the segregation of alleles at each individual locus. This very small Probability indicates that the phenotypes are not in the proportions that we would expect if independent assortment were taking place. In summary.001. we can use the multiplication rule to obtain the probabilities and numbers of progeny inheriting different combinations of phenotypes: The observed and expected numbers of progeny can now be compared by using the chisquare test: Here. then. because the probabilities expected with independent assortment are based on the probabilities expected at the separate loci. we have four expected classes of phenotypes. Rex Arunraj Assistant Prof.BT0203 Genetics and Cytogenetics We are now ready to test for the independent assortment of genes at the two loci. The chi-square tests for segregation at individual loci should always be carried out before testing for independent assortment. is that these genes are not assorting independently and must be linked. Department of Genetic Engineering . so the degrees of freedom equal 4 .

Eg: Mirabilis jalapa Parents : Homozygous red flower RR Gametes: R r X Homozygous White flowers rr Rex Arunraj Assistant Prof. Non – allelic gene interaction Allelic gene interaction The genic interaction occurring between genes located in different rows of the same chromosome or different chromosome is known as non-allelic gene interaction. The genes interaction is of two types namely 1.BT0203 Genetics and Cytogenetics Gene Interaction The expression of a single character by the interaction of more than one pair of alleles is called gene interaction or interaction of genes. The expression of the two alleles (R and r) in the same individual leads to the production of an individual with mixed characters. Department of Genetic Engineering . Example : Mirabilis jalapa When a homozygous red flowered (RR) 4’ o clock plant is crossed which a homozygous white flowered plant (rr) a pink coloured variety is produced (Rr). So the F1 individual has mixture of character of both the parents. This is due to the in complete dominance of the allele R over its allele r. 2. Allelic gene interaction Incomplete dominance In incomplete dominance both alleles of a character express their character in the F1 generation.

None is masked. red and white. generation. Inheritance of coat colour in short horn cattle is another case of codominance Parents (Red) RR F1 generation (Roan) F1 selfed F2 generation RR Red Rr Rr X rr Roan White Rr X (White) rr Rr Rr (Pink Flower) In short horn cattle. This is because r allele also expresses its character in the F1 Rex Arunraj Assistant Prof. the F1 has roan colour having both red & white hairs. there are two colour of hair.BT0203 Genetics and Cytogenetics F1 Generation: Codominance In co-dominance. Department of Genetic Engineering . When red & white are crossed. both alleles of a character an equally dominant and both of them expressed their character in the F1 generation. Red colour is controlled by R & White by r.

The alleles of hypostatic locus (BB.. the inhibited gene is called the hypostatic gene. Dominance Epistasis is is intrallelic or intragenic intergenic Dominant Epistasis (12:3:1) Dominant Epistasis the prevention of the expression.BT0203 Genetics and Cytogenetics Non – allelic gene interaction Epistasis It is the prevention of the expression of one gene by another non-allelic gene. Department of Genetic Engineering . Epistasis means stopping on inhibiting. The allele I prevents the expression of hypostatic gene locus (B / b) and produces white coat color. This is counter part of dominance. of a gene by a dominant non-allelic gene. Bb. Parent : Ii Bb (White) X Ii Bb (White) Rex Arunraj Assistant Prof. bb) express only when two recessive alleles (ii) occur on the epistatic loci iiBB / iiBb produce black and iibb produces brown coat color. The inhibiting gene is called epistatic gene. Inheritance of colour pattern in case of dominant Epistasis. Eg – Colour coats of dogs One gene locus has a dominant epistatic inhibitor allele (I) of coat color pigment.

black. agouti. Eg. Parent Agouti (BBAA) F1 Rex Arunraj Assistant Prof. When F1 was crossed the ratio was 9:3:4. When a homozygous agouti (BBAA) is crossed with homozygous albino (bbaa).BT0203 Genetics and Cytogenetics Gamete IB Gameter IB Ib IB IIBB White Ib IIBb White iB IiBB White ib IiBb White iB ib Ib IIBb White Iibb White IiBb White Iibb White iB IiBB White IiBb White iiBB Black iiBb Black ib IiBb White Iibb White iiBb Black iibb Brown 9:3:3:1 has become 12:3:1 Recessive Epistasis . X Albino(bbaa) Agouti(BbAa) Department of Genetic Engineering . The agouti is wild type.9:3:4 The prevention of the expression of a gene by a recessive non-allelic gene is called recessive epistasis.Coat color in mice The common house mouse occurs in a number of coat colors. the F1 all were agouti. and albino.

The action of these independent genes is complementary. IF both loci have homozygous recessive alleles. Gene C-----. both of them produce identical phenotypes. Duplicate Recessive genes (Complementary genes) – 9:7 Complementary genes may be defined as two or more non-allelic dominant genes interact with one another to produce a character but one gene cannot produce that character in the absence of the other. The chromogen cannot be converted in to anthocyanin In the absence of the enzyme. Eg: Flower colour is Sweet Pea Inheritance of flower colour is sweet pea.Chromogen Rex Arunraj Assistant Prof.BT0203 Genetics and Cytogenetics F1 gametes Gameter BA BA BA BBAA Agouti Ba BBAa Agouti bA BbAA Agouti ba BbAa Agouti Ba bA Ba BBAa Agouti Bbaa Albino BbAa Agouti Bbaa Albino ba bA BbAA Agouti BbAa Agouti bbAA Black bbAa Black ba BbAa Agouti Bbaa Albino bbAa Black bbaa Albino Hence the ratio of 9:3:3:1 become 9:3:4. The red Colour of the flower is due to the presence of a pigment called anthocyanin. Department of Genetic Engineering . The anthocyanin is produced from a colourless substance called chromogen by the action of an enzyme or activator. one producing and flower the other white flower. Two varieties of pea plants. Lathyrutus ordoratus.

Department of Genetic Engineering .BT0203 Genetics and Cytogenetics Gene A ---.Enzyme Chromogen + Enzyme -- anthocyanin (Red) Red flower is produce by the interaction of both dominant gene C and A. C&A cannot give colour independently. Parent White CCaa Gametes F1 generation F1 Selfing CcAa (Red) Ca CcAa (Red) X CcAa (Red) X White ccAA cA Rex Arunraj Assistant Prof.

a new character is expressed. Department of Genetic Engineering . Ex: coat color in Duroc – jersey breed of pigs Sandy – dominant Parent Sandy (SSrr) F1 S_. but when the second dominant gene is added to the first. / R_ X and Sandy (ssRR) Red (SsRr) White ss / rr Rex Arunraj Assistant Prof.BT0203 Genetics and Cytogenetics Gametes Gametes CA CA CA CC AA Red Ca CCAa Red cA CcAA Red ca CcAa Red Ca cA ca Ca CC Aa Red CCaa White CcAa Red Ccaa White cA Cc AA Red CcAa Red ccAA White ccAa White ca Cc Aa Red Ccaa White CcAa White ccaa White Duplicate genes with cumulative effect (Supplementary genes) – 9:6:1 Two independent pair of dominant alleles interact in such a way that each dominant gene produces its effect whether the other is present or not.

X ttdd (Top / Oval) Department of Genetic Engineering . The dominant alleles of both gene loci produce the same phenotype without cumulative effect. both recessive produce oval seed case. or Top/ Oval .D Any one dominant can produce triangular seed case.Seed case in Capsella – Triangular – T.BT0203 Genetics and Cytogenetics F2 gametes Gametes SR SR SR Sr Sr SSRr Red SSrr sR sr sR SsRR Red SsRr Red ssRR Sandy ssRr Sandy sr SsRr Red Ssrr Sandy ssRr Sandy ssrr White SSRR Red Sr SSRr Red Sandy SsRr Red Ssrr Sandy sR SsRR Red sr SsRr Red Hence the Ratio 9:3:3:1 has become 9:6:1 Duplicate Dominant Genes (15:1) Single character controlled by two or more pair of non-allelic genes independently. Parent TTDD (Triangular) Rex Arunraj Assistant Prof. Eg.

Department of Genetic Engineering . Eg. Rex Arunraj Assistant Prof. similarly the white color of feathers of Plymouth rock is caused by the recessive genotype ccii. When both white are crossed the f1 is white. F2 produces white and colored birds in the ratio 13:3.In leghorn fowl the white color of feather is caused by the dominant genotype CCII.BT0203 Genetics and Cytogenetics Gametes F1 Selfing Gametes Gametes TD TD TTDD Triangular Td TTDd Triangular tD TtDD Tringular td TtDd Triangular TD TD td TtDd (Triangular) Td Td TTDd Triangular TTdd Triangular TtDd Tringular Ttdd Triangular tD tD TtDD Triangular TtDd Triangular ttDD Tringular ttDd Triangular td td TtDd Triangular Ttdd Triangular TtDd Tringular ttdd Oval / top Ratio : 15: (Triangular) 1 (Oval) Dominant and Recessive epistasis – 13:3 The dominant alleles of one gene locus (A) in homozygous (AA) or heterozygous (Aa) condition and the homozygous recessive alleles (bb) of another gene locus (B) produce the same phenotype.

BT0203 Genetics and Cytogenetics Parent CCII Gametes F1 F2 gametes Gametes CI CI CI CCII White Ci CCIi White cI CcII White ci CcIi White CI CcIi (White) Ci Ci CCIi White CCii Colored CcIi White Ccii Colored cI ci cI CcII White CcIi White ccII White ccIi White ci CcIi White Ccii colored ccIi White ccii White X ccii ci Hence the ratio is 13:3 Back Cross : (TT X Tt) The F1 individuals obtained in a cross are usually selfed to get the F2 progeny. Such a cross of F1 individual with either of the two parents is known as a backcross. Department of Genetic Engineering . Test Cross : (Tt X tt) (1:1) In such back crosses. when F1 is back crossed to the parent with recessive phenotype. Rex Arunraj Assistant Prof. They can also be crossed with one of the other two parents from which they were derived.

Environmental factors influence Penetrance. These are genes which not only provide certain phenotypic traits but at the same time influence the viability of the organism.BT0203 Genetics and Cytogenetics Penetrance : The percentage of individual’s expressing the character for a particular genotype is called Penetrance. Expressivity The variation in the degree of expression of a particular gene is called Expressivity. If all the individuals expressing the character for a particular genotype. Lethal Genes in mice Yellow coat colour dominant gene Y YY – Lethal effect All yellow individuals are heterozygous (Yy) Two yellow individuals are crossed. Department of Genetic Engineering . 10% people have white eyes even though they contain the BB genes. Ex : Effect of temperature on length of wings in Drosophila . If few individuals do not express the character even though they contain the necessary gene.A particular gene may produce varying degrees of expression in different individuals. the Penetrance is incomplete Penetrance.Expressivity is due to the influence of environmental factor on the genes. Rex Arunraj Assistant Prof.Produce blue eyes – 90% of human beings. the Penetrance is called complete Penetrance. BB. Lethal Genes A lethal gene kills its possessor.

BT0203 Genetics and Cytogenetics Off springs 2Yy, 1yy expected – 1YY : (Lethal) Pleiotropism The production of many characters by a single pair of genes is called Pleiotropism It is the multiple effect of a pair of genes. Intermediate lethal genes Partial effect in heterozygous condition. Eg- Creepers – short, crooked legs - normally they creep. Parent Genotype Gamete: C :creeper Cc c C C CC Normal C Cc Creeper C CC Creeper Cc Dies X creeper Cc 2Yy: 1yy

F1 Selfing Homozygous creeper is missing Balanced Lethal systems Both Homozygotes (Dominant and recessive) die. Rex Arunraj Assistant Prof. Department of Genetic Engineering

BT0203 Genetics and Cytogenetics MULTIPLE ALLELES So for we have observed that a given phenotypic trait is being controlled by two alternate forms of a gene (wild form) and the other being recessive (Mutant form) one bring dominant (Wild form) at the other being recessive (Mutant form). If the wild form mutates to give wise to the mutant form, there why not the wild form should mutate in more than are way to give wise to many mutant forms, on why should not the mutant form, mutate once again to give wise to other mutant forms. So a gene can haw more than two allelomorphs which make a series of multiple alleles. Definition Multiple alleles are a set of three or more allelic form of a gene controlling the same character located on the homologous chromosomes. In other words all the Mutant forms of a single wild type gene constitute a service of multiply alleles. A diploid individual possessor any two alleles of the allelic series and its gametes caries only one allele. Characters of Multiple Alleles. Multiple alleles of a services always occupy the same locus in the chromosome. Because all the alleles of multiple service occupy the seem locus in chromosome, on enclosing over occurs within the same alleles of a multiple service. Multiple alleles always influence the same character. The wild type alleles is nearly always dominant while other mutant alleles may show dominant or not. When two mutant multiple alleles are crosses the phenotype is mutant type all not the wild type Example For Multiple Alleles :

Rex Arunraj Assistant Prof.

Department of Genetic Engineering

BT0203 Genetics and Cytogenetics 1. ABO blood group 2. Rh blood group 3. Coat colour in Rabbit 4. Nature of wing in Drosophila 5. Self Sterility in tobacco. 6. Skin colour in Mice. ABO Blood Grouping in Humans. On the basis of presence or absense of antigens ABO blood groups have been established by K. Landsteiner. Antigen A A B 2 Antigens 4 Blood groups AB

Antingen B


They are called as ABO blood group or Landsteiner blood group. A B AB O group group group group person person person person contain contain contain contain antigen antigen antigen No A B A&B Antigen on “ “ “ RBC “ “ “

Rex Arunraj Assistant Prof.

Department of Genetic Engineering

possible genotypes LO LO LA LO or LA LA Department of Genetic Engineering .exist “ with “ antibody “ A B The three alleles responsible for the inheritance of ABO blood group are.BT0203 Genetics and Cytogenetics Different ABO Blood groups and the antigens & antitrades associated with them. LA LB Responsible “ For “ Synthesis “ of “ Antigen “ A B Lo Absence of Antigen LA LB – produces both Antigen A & B. Blood group Antigen Antibody Antigen for A B AB O A B A and B No Antigen B A No antibody A and B Antigen “ A B cannot “ Co. Different ABO blood grupe on thein genotypes Blood group O A Rex Arunraj Assistant Prof.

1.BT0203 Genetics and Cytogenetics B AB Parent Father LB LO or LB LB LA LB x (A group) Genotype Gamete LA LA LA F1 : LA LB (or) LA LO Ex. 4. 3. Department of Genetic Engineering . AB A group A group B group AB x x x x A B group O group O group O Mother (B group) x LBLO LB LO ( Antigen (or) Agglutinogens present in RBC) (Antibodies (or) Agglutinins present in the serum) A A B AB O + + B + + AB O + + + - Rex Arunraj Assistant Prof. 2.

AB blood group will not agglutinize any other group science no antibodies are present. Compatible donor and recipient Blood group of donor A B AB O (Universal Donor) Blood group of recipient A and AB B and AB AB (Universal Reciepient) O. Department of Genetic Engineering . AB Rex Arunraj Assistant Prof.positive & Rh negative (Rh-) Person Who Donates Blood is called as a Donor Person Who Receives blood is called as a Recipient In transfusion the blood of the donor and the recipient should be compatible while testing the compatibility the reaction between the antigen of the doner’s RBC and the antibodies of the recipient’s plasma alone are taken into consideration Antibodies in blood group A will agglutinize RBC’s of blood group B on vice versa. B. O blood group should be able to agglutinize all other three groups.BT0203 Genetics and Cytogenetics RH bood group There are two groups of human brings namely Rh. A.

replication recombination of cellular DNA.4A. Chromosome provides a definite organization which helps is recombination.BT0203 Genetics and Cytogenetics Unit II Introduction • • • • • • • • • • • • • • • • • • • E. Hence DNA has to be compacted by several orders. . only DNA packed can be efficiently transferred. The chromosome is a compact form of the DNA that readily fits inside the cell. Protects DNA from damage. These are DNA binding proteins which regulate the transcriptions. chromosome DNA is highly stable). DNA with histones will form structures called nucleosomes. Diameter of a typical human cell nucleus is only 10-15 µ. A human cell contains 3x10 9 bp per haploid set of chromosomes. Strasburger in 1875 discovered thread – like structures during cell division. A given region of DNA with its associated proteins is called chromatin. The thickness is 3. Department of Genetic Engineering Rex Arunraj Assistant Prof. Cell cycle – Four phases G1 – Resting phase / pre DNA synthesis phase S – DNA synthesis takes place / DNA synthesis phase inter phase G2 – Resting phase following / Post DNA synthesis phase M – Mitosis division / Mitotic phase Mitosis Duration of different phases not only varies with the organism but with different tissues in the same organism. (Naked DNA is unstable. Each time a cell divides. Packing of DNA into chromosome serves several important functions. Majority of the associated proteins are small basic proteins called histones other proteins are called non-histone proteins. This is true for prokaryotic & Eukaryotic cells & even for viruses.

But all eukaryotic cells have multiple linear chromosomes.BT0203 Genetics and Cytogenetics • The formation of nucleosomes is the first step in a process that allows the DNA to be folded into much compact structure reducing their linear length by 10. • • • • • Ends of linear DNA of the have to be protected from enzyme which will degrade the ends of the DNA. • • Not all cells in eukaryotic organisms are diploid. Even then the DNA has to be compacted. However.g. Department of Genetic Engineering .single circular chromosome In Eukaryotic cell . but code for desirable traits of bacteria. Prokaryotes have one complete copy of their chromosome that is packed into a structure called nucleoid. The majority of eukaryotic cells are diploid i. without which the few daughter molecules remain inter locked after replication. The two copies of a given chromosome are called homologs. They also frequently carry one or smaller independent circular DNAs called plasmids.Multiple linear chromosome. Plasmids like chromosomal DNA are not essential for bacterial growth. Bacteria do not have histones or nucleosomes but have small basic proteins which compact DNA. sperm and egg cells.e. one derived from each parent.000 times. In prokaryotic cell . Haploid cells contain a single copy of each chromosome e. • Circular chromosomes require topoisomerases to separate the daughter molecules after they have replicated. Rex Arunraj Assistant Prof. Some are haploid and some are polyploid. there are numerous examples of prokaryotic cells having multiple chromosomes which are linear. • • • • • Prokaryotic cells have comparatively small genomes. they contain two copies of each chromosome.

Polyploid cells have more than 2 copies of each chromosome. Centromere occurs is the center and forming the equal arms.g. Telocentric Acrocentric Metacentric Terms Chromonemata Centromere on the proximal end Centromere at one end giving a very short arm and a long arm.25u . sub – terminal or median in position. This constriction can be terminal. E.BT0203 Genetics and Cytogenetics • • • Size A chromos is normally measured at mitotic metaphase. The segregation of such a large number of chromosomes is difficult. Megakaryocytes . This clear zone also called as kinetocore which divides the chromosome into 2 arms each one is called a chromosome arm. The position of the centromere varies from chromosome and chromosome providing different shapes. Department of Genetic Engineering . Primary constriction or centromere determines the shape of the chromosome. Submetacentric- Rex Arunraj Assistant Prof.fungi & birds 30 u – plants 3 u – Drosophila 5 u – Man Shape • • • • • Chromosome shape is observed at anaphase. 0. Centromere occurs near the center forming two unequal arms. In extreme case there can be 100 or 1000 copies of each chromosome.128 copies of each chromosome. No matter the number eukaryotic chromosomes are already contained with is a membrane bound organelle called nucleus.

Matrix is enclosed in a sheath or pellicle both matrix and pellicle are material and appear only at metaphase when the nucleolus disappears. If the chromosome breaks. A chromonema represents a chromatid at the early stages of condensation. called chromonemata. A chromosome however cannot fuse at the telomeric ends. Chromosome remains connected with the spherules of the centromere. It is the region of the chromosome to which the spindle fibers of the mitotic phase are attached. Rex Arunraj Assistant Prof. Department of Genetic Engineering . Centromere Consist of granules or spherules.BT0203 Genetics and Cytogenetics During mitotic prophase the chromosomal material becomes visible as very this filaments. Chromonemata form the gene – bearing protions of the non-genetic chromosome. Some species have diffuse centromeres with microtubules attached along the length of the chromosome which are called holocentric chromosomes. The chromonemata is embedded in a achromatic substance called matrix. Chromosomes of most organisms contain only one centromere and are known as monocentric chromosomes. the broken ends fuse due to lack of telomeres. The acentric chromosomes remain in the cytoplasm since they cannot attach to mitotic spindle. since a telomere has a polarity which prevents other segments from joining with it. Chromosomes with two centromeres are called dicentric chromosomes. Chromosomal abnormality Chromosomes may break and fuse forming chromosomes without centromere (acentric chromosomes). Both types of those chromosomal aberations are unstable. Telomeres The chromosome extremities or terminal regions on either side are called telomeres. Chromomeres The chromomeres are bead – like accumulations of chromatin material that are sometimes visible along interphase chromosomes. At metaphase the chromosomes are highly coiled and the chromomeres are no longer visible.

Some species may have special characteristics in their karyotypes. Such a constrictions if present is the distal region of the arm would pinch off a small fragment called satellite. many amphibians have only metacentric chromosomes and plants frequently have heterochromatic regions at the telomeres. gramma = something written).. which is the ratio of the lengths of the long and short arms of the chromosome. secondary constrictions and satellites. The satellite remains attached to rest of the body by a thread of chromatin. A diagrammatic representation of a karyotype (or morphological characteristics of the chromosomes) of a species is called ideogram (Gr. Karyotype and Idiogram All the members of a species of plant or the animal are characterized by a set of chromosomes which have certain constant characteristics. The technique can be improved by determining the socalled centromeric index. the mouse has acrocentric chromosomes. Rex Arunraj Assistant Prof. Generally. in which the pairs of homologues are ordered in a series of decreasing size. their relative size. Department of Genetic Engineering . Sometimes an ideogram is prepared for the diploid set of chromosomes. The term karyotype has been given to the group of characteristics that identifies a particular set of chromosomes. length of the arms. for example. in an ideogram. The individual chromosomes are cut out of the microphotographs and lined up by size with their respective partners. the chromosomes of a haploid set of an organism are ordered in series of decreasing size. These characteristics include the number of chromosomes.BT0203 Genetics and Cytogenetics Secondary Constriction Besides centromere (primary constriction) secondary constrictions can be observed in some chromosomes. and position of the centromere. idios = distinctive. A karyotype of human metaphase chromosomes is obtained from their microphotographs.

since it has a role in the phenotype expression of the genes. Both types of this heterochromatin appear to be connected and together. the following two types of chromatin may be distinguished in the interphase nucleus Euchromatin Portions of chromosomes that stain lightly are only partially condensed. It is thought that in heterochromatin the DNA is tightly packed in the 30nm fibre. DNA is found packed in 3 to 8 nm fibre. this chromatin is termed Euchromatin. Types of heterochromatin In an interphase nucleus. In Euchromatin. A karyotype also suggests primitive or advanced features of an organism. usually there is some condensed chromatin around the nucleolus. and some inside the nucleolus. The Euchromatin is considered genetically active chromatin. called perinucleolar chromatin. It is late replicating (i. it is replicated when the bulk of DNA has already been replicated) and is not transcribed.e. It represents most of the chromatin that disperse after mitosis has completed.. structural genes (i. if any. Heterochromatin In 1928.BT0203 Genetics and Cytogenetics Uses of karyotypes The karyotypes of different species are sometimes compared and similarities in karyotypes are presumed to represent evolutionary relationship. Department of Genetic Engineering .e.. Heterochromatin is characterized by its especially high content of reptititive DNA sequences and contains very few. Euchromatin contains structural genes which replicate and transcribe during G1 and S phase of interphase. called intranucleolar chromatin. Rex Arunraj Assistant Prof. Depending on their staining properties. genes that encode proteins). they are referred to as nucleolar chromatin. Heitz defined heterochromatin as those regions of the chromosome that remain condensed during interphase and early prophase and form the so-called chromocentre.

called dispersed chromatin. it separates from the main component of DNA. called satellite DNA. This most common type of heterochromatin occurs around the centromere. Facultative heterochromatin Rex Arunraj Assistant Prof.BT0203 Genetics and Cytogenetics Dense clumps of deeply staining chromatin often occur in close contact with the inner membrane of the nuclear envelope (i.. The exact significance of constitutive heterochromatin is still unexplained.). the heterochromatic regions can be visualized as regions that stain more strongly or more weakly than the euchromatic regions. In many species. For in the mouse genome. Department of Genetic Engineering .e. Such chromosomes comprising wholly constitutive heterochromatin occur in corn. In the condensed chromosomes. satellite chromosomes or accessory chromosomes and contain very minor biological roles. entire chromosomes become heterochromatic and are called B chromosome. showing the socalled positive or negative heteropyknosis of the chromosomes (Gr. constituting 10 per cent of the total mouse DNA. many phytoparastic insects and salamanders. in the telomeres and in the C-bands of the chromosomes. with the nuclear lamina) and is called condensed peripheral chromatin. Between the peripheral heterochromatin and the nucleolar heterochromatin are regions of lightly staining chromatin. Satellite DNA may have a higher or lower G + C content than the main fraction. Heterochromatin has been further classified into the following types: Constitutive heterochromatin In such a heterochromatin the DNA is permanently inactive and remains in the condensed state throughout the cell cycle. In Drosophila virilis. hetero = different+pyknosis = staining. This DNA is called satellite DNA because upon ultracentrifugation.. constitutive heterochromatin exists around the centromeres and such Pericentromeric heterochromatin occupies 40per cent of the chromosomes. Constitutive heterochromatin contains short repeated sequence of DNA.

histones bind tightly to DNA which is an acid. basic because they are enriched in the amino acids arginine and lysine to a level of about 24 more present. Department of Genetic Engineering . Histone H1 is the least rigidly conserved histone protein. It contains 210 to 220 amino acids and may be represented by a variety of forms even within a single tissue.Barr). These organisms are estimated to have an evolutionary history of at least 600 million years. The best known case is that of the X-chromosomes in the mammalian female. Arginine and lysine at physiological pH are cationic and can interact electrostatically with anionic nucleic acids. the number of Barr bodies is always one less than the number of X chromosomes (i. Thus. the sex chromatin or Barr body (Named after its discoverer. H3 and H4. Barr body contains DNA which is not transcribed and is not found in males. during which they diverged structurally. H1 is present only once per 200 base pairs of DNA (in contrast to rest of the four types of Rex Arunraj Assistant Prof. Canadian cytologist Murrary L. XXX female has two Barr bodies and XXXX female has three Barr bodies). Indeed. being basic.BT0203 Genetics and Cytogenetics Such type of heterochromatin is not permanently maintained in the condensed state. namely H1. in facultative heterochromatin one chromosome of the pair becomes either totally or partially heterochromatic. in humans.e. One of the important discoveries that come from chemical studies is that the primary structures of histones have been highly conserved during evolutionary history. H2A.. Such an evolutionary conservation suggests that the functions of these two histones involve nearly all of their amino acids so that a change in any position is deleterious to the cell. histone H4 of calf and of garden pea contains only two amino acid differences in a protein of 102 total amino acid residues. one of which is active and remains euchromatic. whereas the other is inactive and forms at interphase. There are five types of histones in the eukaryotic chromosomes. histones have had a very similar and crucial role in maintaining the structural and functional integrity of chromatin. For example. Histones Histones are very basic proteins. This conservation of structure suggests that over the eras. H2B.

non-histone proteins display more diversity. Rex Arunraj Assistant Prof. Although for sometime these contractile proteins were thought to be contaminants. About 50 per cent non-histones of chromatin have been found to be structural proteins and include such proteins as action. These non-histones differ even between different tissues of the same organism suggesting that they regulate the activity of specific genes. number of non-histones can vary from 12 to 20. H1 histone is absent in yeast. Saccharomyces cerevisiae. although they must carry out similar enzymatic activities. functioning during chromosome condensation and in the movement of chromosomes during mitosis and meiosis. Non-histones In contrast to the modest population of histones in chromatin. in transcription and in the regulation of transcription. Heterogeneity of these proteins is not conserved in evolution as the histones. and α and β tubulins and myosin. Department of Genetic Engineering . These proteins are not as highly conserved among organisms.BT0203 Genetics and Cytogenetics histones each of which is present twice) and is rather loosely associated with DNA. Histones besides determining the structure of chromatin play a regulatory role in the repression activity of genes. it is now believed that they are vital ingredients of the chromosome. Many of the remaining 50 per cent of non-histones include all the enzymes and factors that are involved in DNA replication. Apparently they are not as important as the histones in maintaining chromosome integrity. In various organisms.

H1. H2B. Histones: small proteins rich in basic amino acid that bind to DNA --> chromatin. Human genome includes 60 ~ 100.000 "genes" occupying <5% of DNA The “C value paradox” – organism complexity does not correlate with genome size Chromosome architecture at the microscopic level DNA packaged into chromosomes. Thus. Eukaryotic DNA is complexed with large amounts of protein to form chromatin. and less elaborately folded & structured. A. eukaryotic chromosomes contain enormous amounts of DNA. H2A. smaller. Prokaryote DNA is usually circular. Nucleosomes 1. H3. similar from one eukaryote to another. Primary coiling of DNA is double helix. which must be packed correctly. is highly extended & tangled during interphase but is condensed into short thick chromosomes during mitosis. Department of Genetic Engineering . and H4 Rex Arunraj Assistant Prof.BT0203 Genetics and Cytogenetics Molecular organization of the eukaryotic genome Three levels of genome organization • • • nuclear genome mitochondrial genome chloroplast genome Genome size: It refers to the amount of DNA in a haploid cell.

B. Higher Levels of DNA Packing 1.000 to 100. and H4). Resembles beads along a string. Looped domain: each loop in the 30-nm fiber contains 20. may control gene expression by limiting access of transcription. Nucleosomes: secondary coiling of DNA around histone core. looped domains coil and fold further compacting chromosome (as in metaphase) Rex Arunraj Assistant Prof. formed from DNA wound around a protein core of 2 copies each of four types of histone (H2A. H2B.BT0203 Genetics and Cytogenetics 2. H3. tertiary coiling of core + linker forms "Solenoid":30-nm chromatin fiber: consists of a tightly wound coil with 6 nucleosomes / turn 2. Department of Genetic Engineering . 2o coiling of DNA around histone core basic unit of DNA packing.00 base pairs.

multiple rounds of replication produce chromatids that remain synapsed together in a haploid number of chromosomes. Rex Arunraj Assistant Prof. not actively transcribed b. homologous chromosomes are tightly paired. Polytene chromosomes have been studied mostly in Drosophila salivary glands. The chromosomes is seen as distinct alternating dark and light bands. Department of Genetic Engineering . The dark bands correspond to more condensed regions of the chromatin.BT0203 Genetics and Cytogenetics Interphase chromatin is much less condensed than mitotic chromatin a. in which chromosomes undergo 10 cycles of replication without separation of the daughter chromosomes. • Can be 1000 times thicker than normal meiotic chromosomes. • Polytene chromosomes are joined together at their centromeres by a proteinaceous structure called a chromocenter. Euchromatin: less condensed. These chromosomes are easy to see with a light microscope because of their large size and precise alignment. Large chromosome consisting of many chromatids formed by rounds of endomitosis following synapsis of the two homologues. • In each polytene chromosome. following synapsis of the two homologues. and the light (interband) regions are less dense regions. is actively transcribed Polytene Chromosome A giant chromosome produced by an endomitotic process in which. This leads to 1024 identical strands of chromatin aligned side by side. Heterochromatin: remains highly condensed.

polytene chromosomes show a banding pattern. big chromosomes and big cells. The largest lampbrush chromosomes are to be found in growing oocytes of newts and salamanders. Interbands also contain genes.000 bp. big enough Rex Arunraj Assistant Prof. 5.BT0203 Genetics and Cytogenetics • When stained. so it is scarcely surprising that they have good lampbrushes. • • In Drosophila. and may contain up to seven genes. Lampbrush chromosomes are exceedingly delicate structures and no further progress beyond the pioneer studies of Flemming (1882) and Ruckert (1892) was possible until a technique could be devised for dissecting them out of their nuclei and examining them in a life-like condition. They have nuclei that are between a third and a half a millimeter in diameter. The chromosomes are greatly elongated diplotene bivalents. • Balbiani ring is a large chromosomal puff. They are about 1 mm in diameter. Department of Genetic Engineering . The best oocytes for lampbrush studies are the ones that make up the bulk of the ovary of a healthy adult female at the time of year when the eggs are actively growing in preparation for ovulation in the following spring. Chromosome puffs are diffused uncoiled regions of the chromosome that are sites of RNA transcription. These animals have big genomes.000 bands have been observed. sometimes reaching lengths of a millimeter or more. separated from the remainder of the nuclear contents. Each band contains an average of 30. Lampbrush Chromosomes The lampbrush type of chromosome is characteristic of growing oocytes in the ovaries of most animals with the exception of mammals and certain insects.

connected by an exceedingly thin thread of the same material (figure). A lampbrush chromosome is a meiotic half bivalent. The entire lampbrush bivalent will therefore have a total of 4 chromatids. and during the period of oogenesis when they are maximally Rex Arunraj Assistant Prof.BT0203 Genetics and Cytogenetics to see with the naked eye. Figure Chromomeres bearing pairs (L) or multiple pairs (LL) of lateral loops: Sister loops of different lengths (L1): polarization of thickness along the lengths of loops (P): Loops consisting of a single unit of polarization (P): Loops consisting of several tandem units of polarization with the same or different directions of polarities (ppp). This means that it must consist of two chromatids. The loops have a thin axis of DNP surrounded by a loose matrix of ribonucleoprotein (RNP). The chromosome appears as a row of granules of deoxyribonucleoprotein (DNP). It is not difficult to isolate these nuclei by hand and it is not much more difficult to remove their nuclear envelopes and spill out their chromosomes. Each chromomere has 2 or some multiple of 2 loops associated with it. Department of Genetic Engineering . Chromomeres are 1/4 to 2um in diameter and spaced 1 – 2 micrometers along the length of the chromosome axis. the chromomeres. The loops are variable in length.

BT0203 Genetics and Cytogenetics developed. Rex Arunraj Assistant Prof. Department of Genetic Engineering . they extend from 5 to 50 micrometers laterally from the chromosome axis. which means that the longest loops in such a case would be 100 micrometers long.

The XY sex-determination system is the sex-determination system found in humans. Other species (including most Drosophila species) use the presence of two X chromosomes to determine femaleness. In this system. One X chromosome gives putative maleness. In the case of humans. Males have two distinct sex chromosomes (XY).BT0203 Genetics and Cytogenetics Sex Determination • • • • Genetic mechanism Metabolically controlled Hormonally controlled Environmentally controlled Genetically Controlled a) Heterogametic Males. Other mammals use several genes on the Y chromosome for that same purpose. Not all male-specific genes are located on the Y chromosome. Some species (including most mammals) have a gene or genes on the Y chromosome that determine maleness. and are called the heterogametic sex. The XY sex determination system was first described independently by Nettie Stevens and Edmund Beecher Wilson in 1905. females have two of the same kind of sex chromosome (XX). XX/X0 sex determination Rex Arunraj Assistant Prof. a single gene (SRY) on the Y chromosome acts as a signal to set the developmental pathway towards maleness. The presence of Y chromosome genes are required for normal male development. most other mammals. some insects (Drosophila) and some plants (Ginkgo). and are called the homogametic sex. Department of Genetic Engineering .

ZZW and ZZWW females can be found. In this system. while females have two (XX). crickets. some fish. Males only have one X chromosome (X0). with a pair of chromosomes (XX) it is a hermaphrodite. This suggests that the W chromosome is essential in female determination in some Rex Arunraj Assistant Prof. females have two copies of the sex chromosome (XX) but males have only one (X0). like the X chromosome in the XY system. The 0 denotes the absence of a second sex chromosome. Department of Genetic Engineering . elegans is male with one sex chromosome (X0). In the ZW system it is the ovum that determines the sex of the offspring. The Z chromosome is larger and has more genes. It is unknown whether the presence of the W chromosome induces female features or the duplication of the Z chromosome induces male ones. unlike mammals. and some other insects use to determine the sex of their offspring. In this variant of the XY system. The letters Z and W are used to distinguish this system from XY system. b) Heterogametic females ZW sex chromosomes The ZW sex-determination system is a system that determines the sex of offspring in birds. referred to as X. and both chromosomes are responsible for gender selection. Its sperm normally contain either one X chromosome or no sex chromosomes at all. Males are the homogametic sex (ZZ). Maternal gametes always contain an X chromosome. examples of Z0. The zero (sometimes. there is only one sex chromosome. It is possible that either condition causes embryonic death. no birds with a double W chromosome (ZWW) or a single Z (Z0) have been discovered. so the sex of the animals' offspring is decided by the male. while females are heterogametic (ZW). and some insects (including butterflies and moths). the letter O) signifies the lack of a second X chromosome. cockroaches. The nematode C. In Lepidoptera (moths and butterflies).BT0203 Genetics and Cytogenetics The X0 sex-determination system is a system that grasshoppers. in contrast to the XY sexdetermination system and the X0 sex-determination system.

BT0203 Genetics and Cytogenetics species (ZZW), but not in others (Z0). In Bombyx mori (the commercial silkworm), the W chromosome carries the female-determining genes. The ZW sex-determination system is found in birds and some insects and other organisms. The ZW sex-determination system is reversed compared to the XY system: females have two different kinds of chromosomes (ZW), and males have two of the same kind of chromosomes (ZZ). c) Haploidy The Haplodiploid sex-determination system determines the sex of the offspring of many Hymenopterans (bees, ants, and wasps), and coleopterans (bark beetles). In this system, sex is determined by the number of sets of chromosomes an individual receives. An offspring formed from the union of a sperm and an egg develops as a female, and an unfertilized egg develops as a male. This means that the males have half the number of chromosomes that a female has, and are haploid. This system produces a number of peculiarities; chief among these is that a male has no father and cannot have sons, but he has a grandfather and can have grandsons. Unfertilized eggs develop into haploid individuals, which are the males. Diploid individuals are generally female but may be sterile males. Thus, if a queen bee mates with one drone, her daughters share ¾ of their genes with each other, not ½ as in the XY and ZW systems. After mating, fertile Hymenopteran females store the sperm in an internal sac called the spermatheca. The mated female controls the release of stored sperm from within the organ: If she releases sperm as an egg passes down the oviduct, the egg is fertilized. Social bees, wasps, and ants can modify sex ratios within colonies to maximize relatedness among members, and to generate a workforce appropriate to surrounding conditions. d) Sex balance theory or genic balance theory Rex Arunraj Assistant Prof. Department of Genetic Engineering

BT0203 Genetics and Cytogenetics

Sex balance theory or genic balance theory states that the X chromosome determines the sex of the individual and that sex is a dosage phenomena, where the ratio of the amount of the X relative to the autosomes determines the sex. In addition, environmental effects can influence the development of the intersex flies. Further studies have shown that sex is ultimately determined by the locus sex-lethal on the X chromosome, though several other loci on the X chromosome and the autosomes are also needed for sex determination.

Environmental sex determination The same chemical plays a unique role in the worm's sexual differentiation. The planktonic, free-swimming Bonellia larvae are initially sexually undifferentiated. Larvae which land on unoccupied sea-floor mature, over the period of years, into adult females. But most larvae come in contact with the bonellin in the skin of an adult female, bonellinrich proboscis (The adult Bonellia female produces a vivid green pigment in its skin, known as bonellin. This chemical, concentrated mostly in the proboscis) and are masculinised by this exposure. The chemical causes these larvae to develop into the tiny males, which cling to the female's body or are sucked inside it by the feeding tube, to spend the remainder of their lives as parasites inside the female's genital sac, producing sperm to fertilize her eggs and reliant on their host for all other needs. The sex of a Green Spoonworm is thus determined by external, environmental factors (the presence or absence of bonellin), not by internal, genetic factors (chromosomes), as is the case with most other sexually-differentiated organisms. This environmental sex determination helps Green Spoonworm populations respond to the availability of burrows. Hormonally controlled sex determination

Rex Arunraj Assistant Prof.

Department of Genetic Engineering

BT0203 Genetics and Cytogenetics Sex reversal in Hen In birds only one gonad of a normal female develops into a functional ovary. The other gonad remains rudimentary. If the functional ovary of a hen is destroyed, the rudimentary gonad develops into a testis. Thus the female sex is reversed into male due to the phenomenon called sex reversal. During embryonic development the XY genotype stimulates the pituitary gland to produce female hormones that cause the gonad of the hen to develop into an ovary. After the development of the ovary the pituitary ceases to produce female hormones due to inhibition of the pituitary by hormones produced by the ovary, thus acting as feed-back system. When the ovary of the hen is removed the steroid cells of the adrenal become active and provoke the development of testis.

Sex Linked Inheritance X-linked Inheritance In animals with XY sex determining mechanism, the X chromosome has many loci, many that have nothing to do with sex as such. The Y is usually smaller and possesses fewer loci that are not the same loci as that on the X chromosome. Thus females that have the same allele at a locus on the X chromosome are homozygous. Different alleles would be heterozygous. Males, because they have only one X, are hemizygous and can have only one allele at a locus. Because of this, one copy of a recessive allele will be expressed in the phenotype in males. In sex-linked inheritance, crosses are not reciprocal. The X-linked pattern is called the criss-cross pattern of inheritance because fathers pass the trait to daughters who pass it on to sons. Rex Arunraj Assistant Prof. Department of Genetic Engineering

Examples of X-linked recessive traits are: 1. 2. 3. If the father has normal vision. Color blindness: Half of the sons of a woman who is heterozygous for color blindness will be color blind. Colorblindness. regardless of sex. however. Duchenne muscular dystrophy. 3. e. D. 4. All the daughters of an affected male will be affected but none of the sons. half of the daughters will also be color blind. Hemizygous males are affected. males transmit their X chromosomes only to daughters and their Y chromosomes only to sons. none of the daughters will be color blind. irrespective of the genotype of their father. Half the children of an affected female will be affected. If the father is color blind. Hemophilia. Females transmit an X chromosome both to sons and daughters. For rare X-linked recessive traits: 1. Twice as many females are affected as males. grandfather—mother—son or maternal uncle—mother—son.g. Homozygous recessive females can arise only from matings in which the father is affected and the mother is heterozygous or homozygous. Note that all of this Rex Arunraj Assistant Prof. A normal vision man and a normal vision woman who is heterozygous for color blindness will produce progeny with a 3:1 ratio of normal vision to color blind. Affected males in a pedigree are related to each other through nonaffected females. Affected males will transmit the gene to all daughters. C. The ratio of affected males-to-females is >>1. 2. who will usually be heterozygous and therefore not affected. but all of the color blindness will be in male progeny. but to no sons. 3. 2. Department of Genetic Engineering .BT0203 Genetics and Cytogenetics Transmission of X-linked genes A. but only homozygous females are affected. B. For rare X-linked dominant traits: 1.

but the good gene on the other X chromosome produces enough of the good clotting enzyme to maintain health. this usually means they must inherit the disease from both parents. Hemophilia: This disorder of blood clotting is X-linked and is common in the males of European royal families because of frequent consanguineous matings within this limited breeding pool. In a carrier. but women are XX. There are actually various types of color blindness involving loss of sensitivity to different colors of light. but this is not the case for X-linked recessive diseases. a gene error on X definitely caused disease in men (who are XY). Pedigrees suggest that Queen Victoria of England was heterozygous for classic hemophilia (hemophilia A). X-linked recessive disorders are more likely to occur than autosomal recessive disorders. Hemophilia B. For autosomal recessive diseases.BT0203 Genetics and Cytogenetics is an oversimplification. because the second gene can pull up the slack. For example. who has only one bad copy. so they have only one copy of any gene on the X chromosome. Holandric or Y-linked Genetic Diseases Rex Arunraj Assistant Prof. Thus. which involves a deficiency of clotting factor VIII. In some recessive diseases. in X-linked recessive hemophilia. a carrier gets a mild form of the disease. whereas all people have 2 copies of each autosome. because men have only one X chromosome. The recessive disease only arises when the male has no good gene on the other chromosome (because they get a Y instead of a second good X). Men have only a single X chromosome. Recessive diseases often occur in genes that produce an enzyme. a female carrier has one bad gene on chromosome X. Department of Genetic Engineering . and maintain health. is also X-linked Recessive means that disease only occurs when a person has two copies of the bad gene. and have two copies of the gene. with a deficiency of clotting factor IX. there is often no disease.

There is evidence of this in rare diseases where the SRY gene is missing. as is a locus concerned with male fertility. however. never to daughters. Since males are hemizygous for Y-linked genes on the nonhomologous part of the Y chromosome. so only men have a Y chromosome. The Y chromosome is very small and contains few genes. There are few genetic diseases related to genes on Y. It is the single gene that sets off the initial cascade of hormone changes that make a person male. but just this gene that is necessary for maleness. Male sex determination: The main Y gene is called the SRY gene. It is not the entire Y chromosome. but with a mutation or deletion of this SRY gene on the Y chromosome. which is the master gene that specifies maleness and male features. C. all will be expressed. antlers in deer. People who are genetically male with XY chromosomes. Sex-limited traits Sex-limited traits are traits that are expressed in one sex. Some examples include milk yield in mammals. and beards in humans. Y-linked genes are transmitted from father to son. And people who are genetically female with XX but also have a tiny piece of the Y chromosome with this gene. Men are XY and women are XX. B. Only men have a Y chromosome and so the Y is only passed from father to son. Department of Genetic Engineering . the gene for maleness is there. There are no essential genes on the nonhomologous portion of the Y.The Y chromosome has a trivially simple inheritance pattern because women are XX and men are XY. will be female despite having most of the Y chromosome. Rex Arunraj Assistant Prof. will become male despite their female-like XX chromosomes. but not in the other. Transmission of Y-linked genes (holandric inheritance) A. Y-linked transmission.BT0203 Genetics and Cytogenetics The Y chromosome is a sex-linked chromosome. The Y chromosome is small and does not contain many genes.

Department of Genetic Engineering . but behaves as a recessive allele in females (must be homozygous to be expressed). Rex Arunraj Assistant Prof. Because of this hormone difference. The male hormone testosterone is needed for full expression of baldness. the allele for baldness behaves as a dominant trait in males (expressed when heterozygous). Male pattern baldness in humans is an example.BT0203 Genetics and Cytogenetics Sex-influenced traits Sex-influenced traits appear in both sexes but more so in one sex than another.

During cell division (Mitosis or Meiosis) each chromosome appears to behave as a unit and therefore it would be expected that genes located on the same chromosome would move to the same pole during cell division. while its recessive allele 'b' determines red flower. it appears that the two dominant gene 'B' (blue colour) and V (elongate pollen grain) have an affinity for each other so that they tend to stay together during the transmission to the progenies. Bateson and Punnet (1905) were the first to report a significant deviation from the expectation due to independent assortment. the progeny exhibited the Rex Arunraj Assistant Prof. the chromosome theory of inheritance. They studied the flower colour in pea. But when F1from the cross Blue Round (BBll) ratio of 1 7 blue elongate (BbLI) blue round (Bbll) Department of Genetic Engineering X Red Elongate (bbLL) Blue Elongate (BbLI) was test crossed with red round (bbll). Sutton expressed this expectation in 1903 while propounding. A dominant gene 'B' produces blue coloured flowers.BT0203 Genetics and Cytogenetics UNIT III LINKAGE AND CROSSING OVER Introduction: Sutton and Bovery proposed that the genes are located on the chromosomes. Bateson and Punnet called this situation as "Coupling Phase". When the F1 from the cross Blue elongate (BBLL) X red round (bbll) was test crossed with red round (bbll). Another dominant gene L governs elongate pollen grains. As a consequence some genes would fail to show independent assortment (segregation) and tend to be inherited together. . where as its recessive allele l produces normal round pollen grains. the progenies showed the ratio of 7 1 1 7 blue elongate (BbLI) blue round (Bbll) red elongate (bbLI) red round (bbll) In the place of expected 1:1:1:1 ratio. Later other workers also proved the same with sufficient evidences.

3) The intensity of the linkage between the two genes on the chromosome is inversely proportional to the distance between the two linked genes on the chromosome. BL and bl in the coupling phase and Bl and bL in the repulsion phase. He concluded that 1) 2) Genes located on the same chrome tend to stay together during inheritance this Genes are arranged in a linear fashion on the chromosome.BT0203 Genetics and Cytogenetics 7 1 red elongate (bbLI) red round (bbll). and explained linkage. but the terms 'coupling' and 'repulsion' phase are widely used still to denote the linkage between two dominant genes an recessive genes respectively. Rex Arunraj Assistant Prof. Morgan worked with gene of Drosophila.e. This situation was referred as "Repulsion Phase". Bateson and Punnet could not give suitable explanation for this situation and suggested that there was a selective multiplication (through mitosis) of some types of gametes i. after meiosis and this gives rise to 7:1:1:7 and 1:7:7:1 ratio instead of 1:1:1:1 test cross ratio. 4) Proposed that coupling and repulsion are the 2 aspects of linkage. tendency is called linkage. so that they tend to stay away from each other in the progeny. It appears as if the two dominant genes (B and L) repel each other in this case. Even though this hypothesis is unstable and later disproved. Department of Genetic Engineering . Morgan’s View on Linkage Later in 1919.

. indicating thereby that these characters are dominant. Bombyx mori crossing-over takes place either very rarely or not at all. Gray. 1.H. Morgan and his co-workers by their investigation on the Drosophila and other organisms have found two types of linkage. In most of the organisms crossing-over takes place both in males and females. F1 progeny had gray bodies and normal long wings (b+vg/ bvg+).. complete linkage and incomplete linkage. Complete linkage in male Drosophila. only two types of progeny (one with gray bodies and vestigial wings. to bvg +/bvg instead of four types of phenotypes were obtained . Morgan mated gray bodied and vestigial winged (b+vg/b+vg) fruit flies with flies having black bodies and normal wings (bvg+/bvg+). Vestigial b+vg/b+vg (b+vg) X Black. test crossed) to double recessive females (bvg/bvg or black vestigial). KINDS OF LINKAGE T.H. Linked genes do not show independent assortment. In this type of linkage genes are closely associated and tend to transmit together.BT0203 Genetics and Cytogenetics LINKAGE: The tendency of two or more genes to stay on the same chromosome is called as linkage. viz. In 1919. When F 1 males (b+vg/ bvg+). Complete Linkage The Complete linkage is the phenomenon in which parental combinations of characters appear together for two or more generations in a continuous and regular fashion. This becomes clear from Morgan’s experimental results from Drosophila. Linkage is the consequence of genes located on the same chromosome. The genes for bent wings (bt) and shaven bristles (svn) of the fourth chromosome mutant of Drosophila melanogaster exhibit complete linkage. b +vg/bvg and the other with black bodies and normal wings. Example. T. Long b+vg/b+vg (bvg+) Department of Genetic Engineering . were backcrossed (i.e. But in male Drosophila and female silkworm. Parents: Gametes: Rex Arunraj Assistant Prof.

long b+vg/ bvg+ X Female Black. vestigial b+vg/ bvg 1Black. When F1 females of the Morgan’s classical cross in Drosophila between gray. the phenotypes of the offspring represent the gametic contribution of the other double heterozygote parent. Long b+vg/bvg+ (b+vg)(bvg+) All Gray. Vestigial X Black. So the genetical analyst can concentrate on one meiosis and forget the other. This sort of exchange of chromosomal segments in between homologous chromosomes is known as crossing over. normal or long (bvg+/bvg+) were test-crossed to double-recessive (bvg/bvg) males. vestigial (b+vg/b+vg) and black. This is in contrast to the situation in an F1 selfing where there are two sets of meiotic divisions to consider one for the F1 male parental gametes and one for the F1 female. all four types of progeny were obtained in following ratio. The linked genes which are widely located in chromosomes and have chances of separation by crossing over are called incompletely linked genes and the phenomenon of their inheritance is called incomplete linkage. Incomplete Linkage The linked genes do not always stay together because homologous non-sister chromatids may exchange segments of varying length with one another during meiotic prophase. Vestigial bvg/bvg (bvg) (Only two types of gametes due to complete linkage and lack of crossing over in male Drosophila) Test cross ratio: 1Gray. Long or 1:1. showing occurrence of crossing-over: Parents: Rex Arunraj Assistant Prof. the use of the testcross is very important. Because one parent (the tester) contributes gametes carrying only recessive alleles.BT0203 Genetics and Cytogenetics F1: Test Cross : Gametes : F1 male Gray. bvg+/bvg Here. Gray. 2. Incomplete Linkage in female Drosophila. Long Department of Genetic Engineering .

3. Crossing over is the exchange of segments between non-sister chromatids of homologous chromosomes. Gray. Long: b+vg+/bvg 4. Vestigial bvg/bvg ↓ (bvg) 83%parental combination showing linkage 17%recombinants due to crossing over bvg+/bvg+ (bvg+) Gametes: (b+vg) (bvg+) = Non-crossovers (b+vg+)(bvg) = Recombinants = 41. Crossing over or recombination occurs at two levels (i) at gross chromosomal level. Gray. called genetic recombination. b+vg/bvg 2. long b+vg/ bvg+ X Male Black. Rex Arunraj Assistant Prof. Characteristics of Crossing Over 1.5% = 8. Vestigial. The crossing over results basically from an exchange of genetic material between non-sister chromatids by break-and-exchange following replication.5% Test cross ratio: 1.BT0203 Genetics and Cytogenetics b+vg/b+vg Gametes: F1 : Test Cross: F1 Female Gray.Morgan.5% = 8. Vestigial. bvg/bvg Crossing Over “The crossing over is a process that produces new combinations (recombinations) of genes by interchanging of corresponding segments between nonsister chromatids of homologous chromosomes” The chromatins resulting from such interchanges of chromosomal parts are known as cross overs. called chromosomal crossing over and (ii) at DNA level. 2.H. Black. A reciprocal exchange of material between homologous chromosomes in heterozygotes is reflected in crossing over. bvg+/bvg 3. Long b+vg/bvg+ ↓ (b+vg) All Gray.5% = 41. Department of Genetic Engineering . The term crossing over was coined by T. Black. Long. In simple.

Types of Crossing Over 1. This type of crossing over is known as germinal or meiotic crossing over. but contains material of two sister chromatids (i.BT0203 Genetics and Cytogenetics 4. This event occurs during the leptotene stage of prophase I and though each chromosome at this stage is visually long and thin thread.7 per cent replication of DNA and 75 per cents synthesis of histones. Mechanism of Meiotic Crossing Over The process of crossing over includes following stages in it.. The meiotic crossing over is universal in its occurrence and is of great genetic significance. Synapsis Rex Arunraj Assistant Prof.e. two DNA molecules plus almost duplicated amount of histones). Somatic or Mitotic Crossing Over When the process of crossing over occurs in the chromosomes of body or somatic cells of an organism during the mitotic cell division it is known as somatic or mitotic crossing over. The frequency of crossing over appears to be closely related to physical distance between genes on chromosome and serves as a tool in constructing genetic maps of chromosomes. called attachment plaques.. attachment of each chromosome by its both ends (telomeres) to the nuclear envelope (i. The chromosomes which tend to undergo recombination due to meiotic crossing over necessarily complete two functions: 1. Germinal or Meiotic Crossing Over Usually the crossing over occurs in germinal cells during the gametogenesis in which the meiotic cell division takes place. viz. The somatic or mitotic crossing over has been reported in the body or somatic cells of Drosophila by Curt Stern. Department of Genetic Engineering . and 2.e..99. crossing over and terminalization. 2. 1. to nuclear lamina) via the specialized structure. duplication chromosomes. synapsis. The somatic crossing over is rare in its occurrence and it has no genetical significance. both of which take place prior to onset of prophase I.

Synapsis often starts when the homologous ends of the two chromosomes are brought together on the nuclear envelope and it continues inward in a zipper-like manner from both ends.BT0203 Genetics and Cytogenetics Synapsis or intimate pairing between the two homologous chromosomes (one maternal and another paternal) is initiated during zygotene stage of prophase I of meiosis I. During the process of crossing over. Thus. Homologues continue to stay in synapsis for days during pachytene stage and chromosomal crossing over occurs due to exchange of chromosomal material between non-sister chromatids of each tetrad. the recombination nodules become visible between synapsis chromosomes. Crossing over by Breakage and Union It is well evident that crossing over occurs in the homologous chromosomes only during the four stranded or tetrad stage. thus. Department of Genetic Engineering . Duplication of Chromosomes The synapsis is followed by duplication of chromosomes (in pachytene). brought into juxtaposition (=being side by side) with its homologous gene on the opposite chromosome. By synapsis each gene is. The resultant pairs of homologous chromosomes are called bivalents. each homologous chromosome of bivalents splits longitudinally and forms two identical sister chromatids which remain held together by an unsplitted centromere. During this stage. In other cases. so it is known as tetrad. producing the same type of alignment. aligning the two homologous chromosomes side by side. At this stage 3 per Rex Arunraj Assistant Prof. The longitudinal splitting of chromosomes is achieved by the separation of already duplicated DNA molecules along with certain chromosomal proteins. two non-sister chromatids first break at the corresponding points due to the activity of a nuclear enzyme known as endonuclease. 2. 3. synapsis may begin in internal regions of the chromosomes and proceed towards the ends. At this stage each bivalent contains four chromatids. so that the two non-sister chromatids cross each other. In pachytene. synapsis is the phase of prolonged and close contact of homologous chromosomes due to attraction between two exactly identical or homologous regions or chromomeres. Then a segment on one side of each break connects with a segment on the opposite side of the break.

The fusion of chromosomal segments with that of opposite one takes place due to the action of an enzyme known as ligase . The crossing over. During diplotene. their transposition and fusion. the non-sister chromatids start to repel each other because the force of synapsis attraction between them decreases.. The number of chiasmata depends on the length of the chromosomes because the longer the chromosome the greater the number of chiasmata. The more apart two genes are located on a chromosome.BT0203 Genetics and Cytogenetics cent synthesis of DNA occurs to fill the gap. Kinds of Crossing Over Rex Arunraj Assistant Prof. The crossing of two chromatids is known as chiasma (Gr. chromosomes detaches from the nuclear envelope and each bivalent is clearly seen to contain four separate chromatids with each pair of sister chromatids linked at their centromeres. The crossing over may take place at several points in one tetrad and may result in the formation of several chiasmata. includes the breaking of chromatid segments. Chiasma frequency or percentage of crossing over. Department of Genetic Engineering . Due to the terminalisation the homologous chromosomes are separated completely. while non-sister chromatids that have crossed over are linked by chiasmata. During diakinesis. thus. chiasma=cross) formation. desynapsis begins. The frequency by which a chiasmata occurs between any two genetic loci has also a characteristic probability. The movement of chiasma is known as terminalisation. In a species each chromosome has a characteristic number of chiasmata. synaptonemal complex dissolves and two homologous chromosomes in a bivalent are pulled away from each other. 4. Terminalisation After the occurrence of process of crossing over. The chromatids separate progressively from the centromere towards the chiasma and the chiasma itself moves in a zipper fashion towards the end of the tetrad. the greater the opportunity for a chiasma to occur between them. The closer two genes are linked lesser the chances for a chiasma occurring between them.

In the double crossing over following two types of chiasma may be formed: (i) Reciprocal chiasma... Single crossing over. of progenies Factors affecting the recombination frequency Rex Arunraj Assistant Prof.. X 100 Total no. In the double crossing over. (ii) Complimentary chiasma. the formation of each chiasma is independent of the other and in it four possible classes of recombination occur. the phenomenon is known as double crossing over. The multiple crossing over occurs rarely.... The single crossing over produces two cross over chromatids and two non-crosses over chromatids.. of recombinant progeny is given by: Frequency of crossing over = ... When crossing over takes place at more than two places in the same chromosome pair then such crossing over is known as multiple crossing over.. 3. The reciprocal chiasma occurs in two strand double crossing over in which out of four chromatids only two are involved in the double crossing over.. Department of Genetic Engineering .... When the chiasma occurs only at one point of the chromosome pair then the crossing over is known as single crossing over..... 1.. the second chiasma restores the order which was changed by the first chiasma and it produces two non-cross over chromatids. Multiple crossing over. The complimentary chiasma occurs when three or four chromatids of the tetrad undergo the crossing over....... In the reciprocal chiasma the same two chromatids are involved in the second chiasma as in the first. Double crossing over. When both the chromatids taking part in the second chiasma are different from those chromatids involved in the first chiasma is known as complimentary chiasma... The complimentary chiasma produces four single cross overs but no non-cross over. Estimation of crossing over-frequency from a test cross: No. 2. Thus..... When the chiasmata occur at two points in the same chromosome....BT0203 Genetics and Cytogenetics According to the number of chiasma following types of crossing over have been described....

Age of female: In general. also show similar effects. In Bajra. Treatment of female.BT0203 Genetics and Cytogenetics The frequency of recombination between two linked genes is affected by several factors. However. Genotypes: Many genes are known to affect the occurrence as well as the rate of recombination. while the removal of metallic ions (through the chelating agents present in the diet) increases the recombination frequency. Chemicals: Injection of females with antibiotics like mytomycin D and actinomycin D promotes recombination. lowest frequency of recombination is observed when the females are cultured at 22 C and increased when temperature is increased Nutrition: The presence of Ca and Mg in food reduces the recombination frequency in Drosophila. Increase in the recombination frequency is noticed in the larvae starved during certain stages of development. As location of genes on the chromosome is fixed. the gene trifton present in the cytoplasm shows reduced recombination frequency. Temperature: In Drosophila. heterogametic sex individuals show relatively lower frequency than the homogametic individuals (XX). Department of Genetic Engineering . Even the Drosophila male showed recombination when treated with X-ray. the frequency of recombination obtained between genes may vary due to sampling error or due to other reasons like: Sex: In Drosophila sp. Thus crossing over between two genes would increase with increase in the distance between them. They are (based on the data from Drosophila melanogaster) Distance between genes: The frequency of cross over between two linked genes is associated with the distance between their location in the chromosome. the frequency of recombination shows progressive decline with the aging in Drosophila. Radiation: Recombination frequency was increased when the females of Drosophila and many organisms when irradiated with X-ray. the frequency of recombination is expected to be constant with little of variation. Plasmogenes: Certain genes present in Drosophila also affect the recombination frequency.with alkylating agents such as ethylmethane sulphonate (EMS). Some genes affect the chromosome pairing and some other genes affect Rex Arunraj Assistant Prof.

Therefore the frequency of crossing over Rex Arunraj Assistant Prof. Crossing over frequency and cross over: Crossing over is responsible for the recombination between linked genes. paracentric inversion reduces recombination between genes located within the inverted segment. or modifying factors. During the process (crossing over) a segment of one of the chromatid becomes attached with the homologous segment of a non-sister chromatid and vise versa. The gametes carrying the recombinant chromatids (which carry the recombination of linked genes) are called cross over types. 3) prevents the recombination when present in homozygous condition. Department of Genetic Engineering . Therefore genes located near the centromere show relatively low recombination frequency than those located away from the centromeres. they often increase the recombination in other chromosomes present in the same cell. Genes for asynapsis is known in many crops. Chromosomal aberrations: In Drosophila. so that each bivalent has four chromatids (four-strand stage). Generally these chromosomal aberrations reduce recombinations in the chromosomes in which they are located. In addition there are evidences to prove that the recombination is also affected by genetic background.BT0203 Genetics and Cytogenetics the synapsis (after pairing). Distance from centromere: Centromeres tend to suppress recombination. Similarly the gametes carrying non cross over chromatids which contain only the parental combination of linked genes called non-cross over types. each chromosome of a bivalent has two chromatids. two recombinant chromatids (involved in the cross over) called cross over chromatids and two original chromatids (not involved in cross over) are produced. The C 3G gene of Drosophila (located on chromosome No. Crossing over takes place during pachytene stage. Translocations also reduce recombination in the vicinity of breaks. A similar effect of inversion and translocation on recombination is also known in plants. while it promotes recombination at heterozygous condition. As a result of a cross over. It is assumed that breaks occur at homologous points followed by reunion of the acentric segments. This produces X like figure at the point of exchange of the chromatic segments is called as 'Chiasma'.

iii) each chiasma does not lead to crossing over and iv) crossing over occurs during diplotene. According to this theory. it appears to be attached with each other through chiasma. this theory is able to explain several phenomena which are not explained by the classical theory. (One plane theory) I. only sister chromatids are associated with each other (where as in classical theory the association is only between non-sister chromatids to produce chiasmata).BT0203 Genetics and Cytogenetics between two genes can be estimated as the frequency of recombinant progeny from a test cross. For example. Chiasmatype Theory: It is proposed by Janssens in 1909 and further expanded by him in 1924. ii) crossing over occurs before diplotene. The available experimental evidences did not support this hypothesis. which is not possible. as per the classical theory three homologous chromosomes are required for a trivalent configuration. They are: plane theory) ii) Chiasmatype theory. This frequency is expressed as percentage and it is the frequency of cross over between the two genes. But on the other hand as per the chiasmatatype theory. Relationship between chiasma and crossing over: Chiasma is the X like configuration observed during diplotene or late pachytene. only two chromosomes are required to pair in the different Rex Arunraj Assistant Prof. iii) through out the entire bivalent. i) a chiasma is formed when chromatid of a chromosome comes to associate with the non-sister chromatid of its homologous chromosomes. iv) each chromatid is the consequence of crossing over event and v) a 1:1 ratio is expected between frequencies of chiasmata and crossing over. Most of the evidences support only the chiasmata type theory. Department of Genetic Engineering i) Classical theory (Two . i) chiasmata are produced due to crossing over. According to this theory. when a homologous chromosome moves away from each other. and it is only a historical significance. II. ii) chiasma formation is the cause of crossing over. Two divergent theories have been proposed to explain the relationship between chiasma and crossing over. Further. Classical Theory: Proposed by Sharp in 1934 also called as two-plane theory. This is also called as one-plane theory.

While according to Holiday it occurs in the strands having the same polarity. there is some DNA synthesis associated with crossing over and this DNA synthesis most likely involved in the repairing of chromatid breaks. Breakage and reunion Copy choice theory 1. 2. i. the most important evidence against the chiasmatatype theory comes from the male Drosophila where chiasmata are present but there is no crossing over. Rex Arunraj Assistant Prof. Most likely the chiasmata in male Drosophila are produced due to simple association between homologous chromosomes. Holiday model is relatively simpler and more attractive. Hence. several models have been proposed and these models are called as hybrid DNA models.e. This is well proved by 1) by isotope labeling that revealed each cross over chromatids consisted of segments from two non sister chromatids involved in crossing over.e. Whitehouse proposed that single strand breaks occurs in the strands having opposite polarity. However. The segments of two non-sister chromatids reunite to produce cross over chromatids. AH these hybrid DNA models are the modifications of either the White House model (1963) or the Holiday model (1964).BT0203 Genetics and Cytogenetics segments to yield the same trivalent configuration. Breakage and reunion: This theory was put forth by Darlington in 1932 and explained in simple words as. break occurs at identical points in one of the two chromatids of each of the two homologues forming bivalent . These two models differ in one important aspect i. 2) as a rule. In maize. chiasmata theory is still universally accepted one.. "after synapsis. Theories on crossing over: Several models have been proposed to explain the molecular mechanism of crossing over and these models may be grouped into two broad classes: 1. they are not chiasmata in real sense. Based on this theory. Department of Genetic Engineering . there is a close correlation between the relative length of chromosomes determined on the basis of recombination data and those estimated from the number of chiasmata However.

or at least replication of the segment involved in crossing over. Consequently the newly produced DNA molecule is a recombinant.BT0203 Genetics and Cytogenetics 2. According to this theory. the new copies of genes present in a segment of one chromosome may sometimes become joined with those of the neighbouring segment of the homologous chromosome. But this is contrary to the fact in eukaryotic chromosome replication occurs before synapsis and DNA replication is universally semi-conservative. Therefore the copychoice model is not acceptable. chromosome replication. during DNA replication. must occur after synapsis. This theory requires two important pre-requisites viz.. however. Rex Arunraj Assistant Prof. hence this postulate appears to be unrealistic. 3. in some prokaryotes the DNA replication appears to be copy-choice type. which is contrary to the known facts. i) ii) chromosome replication takes place after synapsis and DNA replication is conservative. Department of Genetic Engineering . a DNA molecule serves as a template (for the DNA molecule to synthesis new DNA molecule) upto a certain distance after which the homologous DNA molecule is used as the template. which gives rise to cross over or recombinant chromatids.Duplication theory: This theory is mostly based on the mechanism of crossing over proposed by Belling in 1933. Copy-choice Model In 1955 Laderberg proposed a modification in Belling's hypothesis to explain the same unusual features of recombination in bacteria. He postulated that i) genes present in a chromosome are the first to be replicated ii) they are subsequently connected with each other through the synthesis of the remaining parts (other than genes of the chromosome) and iii) The homologous chromosome are likely to be coiled with each other so that the newly produced copies of genes present in a segment of one chromosome would be adjacent to those of the neighbouring segment of the homologous chromosome . As a result. This modification is commonly known as copy-choice theory. According to this theory.

BT0203 Genetics and Cytogenetics 4. A map unit is defined as that distance in a chromosome which permits one percent of Rex Arunraj Assistant Prof. The two nicks present in the molecules are sealed by DNA ligase.Such a diagrammatic representation depicting the linked genes and the recombination frequencies between them is known as linkage map or genetic map or chromosome map. Each recombinant molecule has a nick which is finally sealed by DNA ligase. this yields two recombinant DNA molecules. LNKAGE GROUPS AND LINKAGE MAPPING All the linked genes form a linkage group. Such hybrid DNA molecules have been actually isolated from bacteria and photographed with the help of electron microscope. These percentages of recombination frequencies are used as map units for construction of linkage maps. The free strands now pair with the intact strands of the homologous DNA molecule during the unwinding of chromatids. An endonuclease now induces nicks in the two intact (not cut earlier) strands of the hybrid molecules. Then the hybrid DNA molecule undergoes reorientation to form X-shaped figure. ii) the order or sequence of these genes in the chromosomes. The two strands of each DNA molecule separate from each other upto certain distance from the point of 'nick'. Most of the evidences are in favour of this model since almost all the enzymes and proteins involved in the process are known to occur in bacteria. In such representation. Cross.over theory: According to this theory. Thus for preparing a chromosome map the following two information are required: i) the frequency of recombination between linked genes. The genes that are linked may be represented on a straight line in the same order in which they are normally present in the concerned chromosome. This produces a "hybrid DNA molecule". the distance between any two neighbouring genes is proportional to the frequency of recombination between them. one end of this X rotates to 180°. an endonuclease produces single-strand nicks at identical points in the two homologous DNA molecules in strands having the same polarity. Department of Genetic Engineering . As we have already seen the recombination frequency between any two genes can be estimated from appropriate test crosses or F2 progenies.

Since the map units are the recombination frequencies between two linked genes. they are likely to be influenced by several factors. mm etc. The data from a test cross involving three genes will provide the information on the order of the three genes in the chromosome as well as their recombination frequencies between them. If the crossing over between the genes in test cross is more than 20% the linkage maps would not be very reliable. It should be noted that a map unit is an imaginary distance and it does not represent the actual distance between two linked genes in the chromosome. Department of Genetic Engineering .BT0203 Genetics and Cytogenetics recombination between the linked genes. In general the relative length of linkage group of a species correspond closely with the relative lengths of the chromosomes in which they are located. Sometimes the Rex Arunraj Assistant Prof. Therefore a map unit does not have a unit of measurement like A. These linkage maps provide the information on i) the genes that are linked together and ii) the frequencies of recombination that may be expected between them. In such studies it is desirable to include only those genes show less than 20% preferably 10% or less recombination with each other to avoid confusion due to double and triple cross overs.. To begin with a three point test cross involving any two of these three linked genes and a new gene expected to be linked with them is studied to map the new gene. The number of linked group in a species is a rule. equal to its gametic chromosome number (n). DETERMINATION OF THE SEQUENCE OF LINKED GENES: The sequence of linked genes can be determined by studying the test crosses for three genes at a time. Each linkage group of a species is to be assigned to a specific chromosome of that species with the help of chromosomal aberrations. In this manner each new gene is mapped by ordering it in three-point test cross with two already mapped genes. u. A map unit is also referred as Morgan (1 centiMorgan = 1 map unit). In addition the number of test cross progeny should be sufficiently large to yield reliable recombination frequencies and to avoid the effects of sampling error.

So the comparison between parental and double cross over shows that only the gene located in the middle is shifted. However. .. Rex Arunraj Assistant Prof. Hence.BT0203 Genetics and Cytogenetics distance between any two genes in a linkage group may exceed 50% or even 100 %.In this case the double cross over produced will be between 'C and 'Sh' and 'Sh' and 'Wx'. parental types are CShWx and cshwx and the double cross over types are CshWx and cShwx. Department of Genetic Engineering . the parental types are CWxSh and cwxsh and the double cross overs are CWxsh and cwxSh. But there is a progressive decline in the frequency of observed recombination for every additional map unit distance beyond 20 map units. ALTERNATE PROCEDURE TO FIND THE ORDER OF THE GENES: An alternate procedure for determining the gene frequency involves the comparison of recombination frequencies. therefore these double cross over frequencies should also be included to estimate recombination frequencies between 'c' and 'sh' as well as between 'sh' and 'wx'. But in the given table. In a double crossing over simultaneous cross over occurs on both sides of the genes located in the middle.. In the figure the gene 'sh' is located between 'C and 'Wx. Therefore the gene 'sh' must be located between 'c' and 'wx'. but that does not mean that they show recombination beyond 50%. the allele 'sh' have been shifted their position in the double cross overs as compared to parental types. there is a 1:1 correspondence between map distance and the observed recombinant frequency upto 20 map units. which are linked. If the genes 'c\ 'sh' and 'wx' are placed in the order of c-sh-wx single cross over between 'c' and 'sh 'which yield the cross over gametes viz. DETERMINATION OF GENE SEQUENCE: Now using this data the gene sequence can be obtained by comparing the genotypes of the parental and double cross over types. double cross over viz.e.. i. the recombination between two linked genes cannot exceed 50% which is the frequency expected in independent assortment. CshWx and cShwx are also produced due to simultaneous crossing over between genes 'c' and 'sh' and 'sh' and 'wx'. This property of the double cross overs is used for determining the sequences of the genes. Cshwx and cShWx. In addition to the above frequencies between 'c' and 'Sh' and between 'sh' and 'wx'.

9 18. ■ COEFFICIENT OF INTERFERRENCE: As a rule the observed frequencies of double cross overs are less than the expected values... of Double cross overs Similarly single cross over between 'sh' and 'wx' will produce the cross over types viz. this is called as interference. It may be expected that the intensity of interference would decrease as the point of Rex Arunraj Assistant Prof. two genes located on either sides of the gene 'sh'. 3.1 + Double cross overs Now let us consider that crossing over are between 'c' and 'wx' i... In such case the crossing over between *c' and 'sh' and 'sh' and 'wx' would produce recombination between 'c' and 'wx'. Therefore the recombination frequency between C and wx = frequency of c and sh + frequency of sh and wx (It should be noted that double cross over does not lead to recombination between 'c' and 'wx' although two events of crossing over occur between them). . Thus. CShwx and cshWx and the sum of these frequencies will be their recombination between 'sh' and 'wx' i. i) ii) iii) a map distance is the frequency of recombination upto 20 map units linked genes would show independent segregation if they are more than 80 map units apart and maximum recombination observed between two linked gene is 50%.7 1. . 1.. 9.8 . Department of Genetic Engineering ..2 8.e.5 +freq. CShwx cshWx Total ....e.. .. This is because" the occurrence of crossing over in one region of a chromosome interferes with its occurrence in the neighbouring segment. A map distance of around 90 units is expected to show to 50% recombination.BT0203 Genetics and Cytogenetics Therefore the frequency of recombination between 'C and 'Sh' will be the sum of the frequencies of Cwxsh cWxSh Total .

BT0203 Genetics and Cytogenetics second crossing over becomes farther from that of the first one. The intensity of interference may be estimated as coefficient of interference = 1 coefficient of coincidence. Department of Genetic Engineering . Interference=%of observed double cross overs / %of expected cross overs Rex Arunraj Assistant Prof. Therefore the coefficient of coincidence will be lower when the concerned genes are located close to each other.

Genes a and b may therefore either be on different chromosomes or be very far apart on the same chromosome. so we will place them in different linkage groups with the understanding that they may or may not be on the same chromosome: Linkage group 1 a Linkage group 2 The recombination frequency between a and c is 50%. = 30 m. and obtained the following recombination frequencies: Gene loci in testcross a and b a and c a and d b and c b and d c and d Recombination frequency (%) 50 50 50 20 10 28 We can begin constructing a genetic map for these genes by considering the recombination frequencies for each pair of genes. and d. we must consult the c-to-d distance. The recombination frequency between a and b is 50%. If gene d is 10 map units to the left of gene b. + 10 m. with a recombination frequency of 10%. are in different linkage groups. Department of Genetic Engineering . which is the recombination frequency expected with independent assortment.u. c. too.u.BT0203 Genetics and Cytogenetics Constructing a Genetic Map with Two-Point Testcrosses Genetic maps can be constructed by conducting a series of testcrosses between pairs of genes and examining the recombination frequencies between them.u. The recombination frequency between b and c is 20%. Suppose that we carried out a series of two-point crosses for four genes. indicating that these genes belong to different linkage groups. This distance will be only approximate because any double crossovers between the two genes will be missed Rex Arunraj Assistant Prof. so these genes are linked and separated by 20 map units: Linkage group 1 b a Linkage group 2 b 20 c mu The recombination frequency between a and d is 50%. a. whereas genes b and d are linked. b. A testcross between two genes is called a two-point testcross or a two-point cross for short. then the distance between d and c should be 20 m. indicating that they. To decide whether gene d is 10 map units to the left or right of gene b.

on the other hand.e. If. so gene d must lie to the left of gene b.u..u. gene d lies to the right of gene b. causing an underestimation of the true recombination frequency. . Notice that the sum of the recombination between d and b (10%) and between b and c (20%) is greater than the recombination between d and c (28%). The recombination frequency between c and d is 28%. we can distinguish between these two possibilities. = 10 m. map units—are approximately additive. By examining the recombination frequency between c and d.BT0203 Genetics and Cytogenetics and the recombination frequency will be underestimated. Department of Genetic Engineering .10 m. then the distance between gene d and c will be much shorter.u. approximately 20 m. (This is what was meant by saying that recombination rates—i. The genetic map of these genes is now complete: Linkage group 1 a Linkage group 2 d 10 mu b 20 mu c Rex Arunraj Assistant Prof.) This discrepancy arises because double crossovers between the two outer genes go undetected.

BT0203 Genetics and Cytogenetics Linkage and Recombination between Three Genes Genetic maps can be constructed from a series of testcrosses for pairs of genes. In progeny that result from a double crossover. because numerous two-point crosses must be carried out to establish the order of the genes and because double crossovers are missed. In each type of crossover.c ). Consider what happens when crossing over takes place among three hypothetical linked genes. but the middle allele is different. A more efficient mapping technique is a testcross for three genes (a three-point testcross. Gene Mapping with the Three-Point Testcross Rex Arunraj Assistant Prof. or threepoint cross). the outer two alleles are the same as in the nonrecombinants. Department of Genetic Engineering . but this approach is not particularly efficient. Notice that the genes are in the coupling configuration. Three types of crossover events can take place between these three genes: two types of single crossovers (Figure ) and a double crossover (Figure). that is. the order of the three genes can be established in a single set of progeny and some double crossovers can usually be detected. providing more accurate map distances. two of the resulting chromosomes are recombinants and two are nonrecombinants. in the recombinant chromosomes resulting from the double crossover. This result provides us with an important clue about the order of the genes.b---. Figure illustrates a pair of homologous chromosomes from an individual that is heterozygous at three loci (AaBbCc). all the dominant alleles are on one chromosome ( A----.With a three-point cross.C ) and all the recessive alleles are on the other chromosome ( a---. Notice that.B----. only the middle allele should differ from the alleles present in the nonrecombinant progeny.

no crossing over takes place in males. although the testcross can also be done with genes in repulsion. we cross the F1 heterozygotes with flies that are homozygous for all three recessive mutations. a cross between a fly heterozygous at all three loci and a fly homozygous for recessive alleles at all three loci. All three mutations are linked and located on the third chromosome. we will consider three recessive mutations in the fruit fly Drosophila melanogaster. between st+ and ss+. In the three-point testcross. we could just as easily say that there are 10 m. e. it makes no difference whether the heterozygous parent in the testcross is male or female (provided that the genes are autosomal) but. We will refer to these three loci as st. Rex Arunraj Assistant Prof. e+. when we say that there are 10 m.u. we mean that there are 10 m. For two classes of Department of Genetic Engineering . Additionally. between st and ss. st+ ss+ st ss e+ e fema le X st ss st ss mal e e e produced cross are FIGURE. To map these genes. So we mate female F1 flies that are heterozygous for all three traits with male flies that are homozygous for all the recessive traits: The progeny from this listed in each locus. and spineless (ss)—that is. e.u. we need to determine their order on the chromosome and the genetic distances between them. the presence of small bristles—is recessive to normal bristles (ss+).BT0203 Genetics and Cytogenetics To examine gene mapping with a three-point testcross. So. In this species. scarlet eyes (st) are recessive to red eyes (st+). in Drosophila. the alleles in these heterozygotes are in coupling configuration (because all the wild-type dominant alleles were inherited from one parent and all the recessive mutations from the other parent). First. In many organisms. and ss) or the dominant alleles (st+. the heterozygous flies in our testcross must be female. ebony body color (e) is recessive to gray body color ( e+). and ss. and ss+) may be present at each locus. we might cross a stock of flies that are homozygous for normal alleles at all three loci with flies that are homozygous for recessive alleles at all three loci: P st+ ss+ st+ ss+ e+ X e+ st st e e ss ss F 1 st+ e+ ss+ The order of the genes has st e ss been arbitrarily assigned because at this point we do not know which is the middle gene. between the loci at which these mutations occur. To produce flies heterozygous for all three loci. we must set up a three-point testcross.u. but keep in mind that either recessive alleles (st. Because crossing over in the heterozygous parent is essential for determining recombination frequencies.

Because chromosomes contributed by the homozygous parent carry only recessive alleles. one or more of the phenotypes may be missing if the number of progeny is limited.) Among the progeny of the testcross in Figure. In Figure aove. e. therefore. displaying the dominant trait. First. Determining the gene order The first task in mapping the genes is to determine their order on the chromosome. (Even if crossing over takes place in every meiosis. determine which progeny are the nonrecombinants— they will be the two most-numerous classes of progeny. we arbitrarily listed the loci in the order st. with or without crossing over. but we had no way of knowing which of the three loci was between the other two. so crossing over can be detected. As a shortcut. in some threepoint crosses. ss. The information that we need for mapping. the most numerous are those with all three dominant traits st+ e+ ss+ and those with all three recessive traits st Rex Arunraj Assistant Prof. With two classes of progeny possible for each of the three loci. Nevertheless. displaying the recessive trait. we need information about where and how often crossing over has occurred. which are the alleles inherited from the heterozygous parent. comes entirely from the gametes produced by the heterozygous parent. the nonrecombinants will comprise at least 50% of the progeny. the two alleles at each locus are the same. e ss Department of Genetic Engineering .e---. all gametes from this parent have a chromosome with three recessive alleles ( --st--. In contrast. the heterozygous parent has different alleles on its two chromosomes.BT0203 Genetics and Cytogenetics progeny are produced: progeny that are heterozygous. as we will and so crossing over will have no effect on the types of gametes produced. In this example. listing instead only the alleles expressed in the phenotype (as shown in Figure). there will be 2 3 = 8 classes of phenotypes possible in the progeny. and progeny that are homozygous. we usually do not write out the complete genotypes of the testcross progeny. To map the genes. the absence of a particular class can provide important information about which combination of traits is least frequent and ultimately the order of the genes. whatever alleles are present on the chromosome contributed by the heterozygous parent will be expressed in the progeny.). all eight phenotypic classes are present but. We can now identify the middle locus by examining the double-crossover progeny. In the homozygous recessive parent.

Again the only trait that differs is the one for bristles. Determining the locations of crossovers Rex Arunraj Assistant Prof. if they differ in two loci. and normal bristles ( st e ss+ ). The three possible gene orders and the types of progeny produced by their double crossovers are: The only gene order that produces chromosomes with alleles for the traits observed in the double crossovers (st+ e+ ss and st e ss+) is the third one. where the locus for bristle shape lies in the middle. we can draw the chromosomes of the heterozygous parent with all three possible gene orders and then see if a double crossover produces the combination of genes observed in the doublecrossover progeny. ebony body. but the nonrecombinants have an allele for normal bristles (ss+). gray body. The phenotypes of the progeny are expressions of the alleles inherited from the heterozygous parent. With a little it’s possible to quickly determine which locus is in the middle without writing out all the gene orders. it must lie in the middle. whereas the double crossovers have an allele for spineless bristles (ss). identify the double-crossover progeny. Recall that.) must be the correct sequence of genes on the chromosome. To determine which gene is in the middle. Therefore. If we compare the nonrecombinant progeny with double-crossover progeny. Department of Genetic Engineering . Let’s compare the alleles in the double-crossover progeny st+ e+ ss with those in the nonrecombinant progeny st+ e+ ss+ . and spineless bristles( st+ e+ ss ) and progeny with scarlet eyes. We see that both have an allele for red eyes ( st+) and both have an allele for gray body (e+). they should differ only in alleles of the middle locus. this order (-. or the bristle locus could be in the middle ( st ss e ).st—e—ss-. Don’t forget that the nonrecombinants and the double crossovers should differ only at one locus. The least-common progeny among those listed in Figure are progeny with red eyes.BT0203 Genetics and Cytogenetics Next. only the alleles at the middle locus differed from the nonrecombinants. the body-color locus could be in the middle ( st e ss ).). when we looked at the results of double crossovers.We would obtain the same results if we compared the other class of double-crossover progeny ( --st ---e –ss+ --) with other nonrecombinant progeny ( because the probability of a double crossover is always less than the probability of a single crossover. Because the bristle locus is the only one that differs. Three orders of genes are possible: the eye-color locus could be in the middle ( e st ss ). These should always be the two leastnumerous phenotypes. so they are the double-crossover progeny.e-. the wrong classes of progeny are being compared.

Department of Genetic Engineering . This same crossover also produces the st ss+ e+ progeny. This same method can be used to determine the location of crossing over in the other two types of singlecrossover progeny. To determine the map distances accurately. consider progeny with chromosome st+ ss e . just as we did for the double crossovers. Calculating the recombination frequencies Next. we have already identified two classes as nonrecombinants ( st+ ss+ e+ and st ss e ) and two classes as double crossovers ( st+ ss e+ and st ss+ e ). Some of the alleles in the singlecrossover progeny are derived from one of the original (nonrecombinant) chromosomes of the heterozygous parent. To determine where the crossovers took place in these progeny. Crossing between ss and e produces st+ ss+ e and st ss e+ chromosomes (figure). The first allele (st+) came from the nonrecombinant chromosome st+ ss+ e+ and the other two alleles (ss and e) must have come from the other nonrecombinant chromosome st ss e through crossing over(figure). dividing this number by the total number of progeny from the cross. which are based on the frequencies of recombination. and two underwent single crossovers between ss and e. we must include all crossovers (both single and double) that take place between two genes. The position of the switch indicates where the crossover event took place. we should rewrite the phenotypes of the testcross progeny with the loci in the correct order so that we can determine where crossovers have taken place ( FIGURE ). The other four classes include progeny that resulted from a chromosome that underwent a single crossover: two underwent single crossovers between st and ss. For example. compare the alleles found in the single-crossover progeny with those found in the nonrecombinants.BT0203 Genetics and Cytogenetics When we know the correct order of the loci on the chromosome. and multiplying the number obtained by 100%. Recombination frequency is calculated by adding up all of the recombinant progeny. Among the eight classes of progeny. Recombinant progeny that possess a chromosome that underwent crossing over between the eye-color locus ( st) and the bristle locus (ss) include the single crossovers ( st+ / ss e and st / ss+ e+ ) and the two double crossovers ( st+ / ss / e+ and Rex Arunraj Assistant Prof. but at some place there is a switch (due to crossing over) and the remaining alleles are derived from the homologous nonrecombinant chromosome. we can determine the map distances.

2 m.).u.) add up to the distance between st and e (26. 26. We can now use the map distances to draw a map of the three genes on the chromosome.u. The recombinant progeny that possess a crossover between ss and e are the single crossovers st+ ss+ / e and st ss / e+ .8 mu st ss e 12. so the recombination frequency between ss and st is: st–ss recombination frequency = (50+52+5+ 3)/755 X 100 = 14.8% Notice that the distances between st and ss (14.8 m.2 m.u. The recombination frequency is: ss–e recombination frequency = (43+41+5+3)/755 X 100 = 12.BT0203 Genetics and Cytogenetics st / ss+ / e ). Rex Arunraj Assistant Prof.2% Thus.2 mu Interference 14.) and between ss and e (12. we must add the double crossovers twice to determine the recombination frequency between these two loci: st– e recombination frequency = (50+52+43+41+(2X5)+(2X3)) / 755 X100 = 26.6 m. which is the ratio of observed double crossovers to expected double crossovers.6 mu The degree to which one crossover interferes with additional crossovers in the same region is termed the interference.6% The distance between the st and ss loci can be expressed as 14. These progeny include those with a single crossover between st and ss. the genetic distance between ss and e is 12.u. The map distance between the bristle locus (ss) and the body locus (e) is determined in the same manner. Department of Genetic Engineering . Add up all the progeny with crossovers between the two loci. Because the double crossovers have two crossovers between st and e.u. Finally. calculate the genetic distance between the outer two loci. those with a single crossover between ss and e.6 m. st and e. Coefficient of coincidence. There are a total of 755 progeny. and the double crossovers st+ / ss / e+ and st / ss+ / e . and the double crossovers ( st+ / ss / e+ and st / ss+ / e ).

In multicellular organisms the germ cells are distinct cells... The first scientific study of mutations was started in 1910. In the drosophila. guinea pigs and man. Oenothera lamarckiana. mutation causes white and pink eyes. mice and other rodents. they are not directed according to the requirements of the organism. so a mutation can be regarded as a change in the DNA sequence which is reflected in the change of sequence of corresponding RNA or protein molecules. HISTORICAL BACKGROUND Hugo de vries was the first hybridist who used the term “mutation” to describe the heritable phenotypic changes of the evening primrose. The discovery of white eyed mutants in Drosophila was followed by Morgan and his co-workers and other geneticists. and vestigial wings.) A unicellular organism is more subjected to environmental onslaughts since it is at the same time a somatic or germ cell. Drosophila melanogaster and reported white eyed male individuals among red eyed male individuals. drosophila. rabbits. Department of Genetic Engineering . Many mutations described by de vries in Oenothera lamarckiana. however. Rex Arunraj Assistant Prof. Mutation occurs in a random manner. Such a change may involve only one base/base pair or more than one base pair of DNA. they can be induced in the laboratory either by radiations. Mutants occur frequently in the nature and have been reported in many organisms. are known to be due to variation in chromosome number or ploidy and chromosomal aberrations (viz gross mutations).. when Morgan and his work was in fruit fly. Most mutations occur spontaneously by the environmental effect. physical factors or chemicals (called mutagens.BT0203 Genetics and Cytogenetics Unit-IV Mutation A Gene mutation is abrupt inheritable qualitative or quantitative change in the genetic material of an organism. Consequently about 500 mutants of Drosophila have been reported by geneticists all over the world.g. Mutation has a significant role to play in the origin of species or evolution. black and yellow body colours. Since in most organisms genes are segments of DNA molecule. rats. and are relatively protected from the environment. ie. e.

KINDS OF MUTATIONS 1. The mutations occurring in non-reproductive body cells are known as somatic mutations. skin pigmentation and several somatic malformations.. Various genetical diseases of human beings such as haemophilia. Thus the amino acid sequence and hence the structure and properties of the enzyme formed will be changed. Examples of somatic mutation have been reported in Oenothera lamarckiana (Hugo de Vries) and several other cases including man. Department of Genetic Engineering . Classification of Mutation According to Type of Cells According to their occurrence in somatic and germinal cells following types of mutations have been classified: Somatic Mutations. a somatic mutation occurs early during embryonic life. however. In man the mutations cause variation in hair colour. eye colour. form other examples for mutation in human beings. How does a mutation act? Any change in sequence of nucleotides in the DNA will result in the corresponding change in the nucleotide sequence of mRNA. This may result in alignment of different RNA molecules on mRNA (during protein synthesis). the mutant cells may constitute a large proportion of body cells and the animal body may be a mosaic for different type of cells. since only single cells and their daughter cells are involved. The genetical and evolutionary consequences body cells are insignificant. Rex Arunraj Assistant Prof. In consequence a mutant phenotype makes its expression. If. white and brown coats. colour blindness.BT0203 Genetics and Cytogenetics In rodents the mutations are responsible for black. This defective enzyme structural protein may adversely affect the trait controlled by the protein. unilateral retinoblastoma and heterochromia of the iris. phenylketonuria etc. In man somatic mutation causes several fatal diseases such as paraoxysmal nocturnal hemoglobinura. Somatic mutations have been often related with malignant (cancerous) growth.

The mutations which arise from the insertion or deletion of individual nucleotides and cause the rest of the message downstream of the mutation to be read out phase. When heritable alterations occur in a very small segment of DNA molecule. i.. The mutations occurring in gamete cells (e. are called frameshift mutations. A point mutation in which a nucleotide of a triplet is Rex Arunraj Assistant Prof. At replication. The insertion mutations can be artificially induced by certain chemical substances called mutagens such as acridine dye and proflavin. Substitution mutation. 3. 1. Deletion mutations have been frequently reported in some bacteriophages. Such mutations are heritable and of immense genetical significance. They result in the production of an incorrect. Insertion or addition mutation. due to which the death of the cell may occur. The point mutation which is caused due to loss or deletion of some portion (single nucleotide pair) in a triplet codon of a cistron or gene is called deletion mutation. A proflavin molecule. Deletion mutations. thereby stretching the strand lengthwise. 2. Department of Genetic Engineering . inactive protein. this situation would allow the insertion of an extra nucleotide in the complementary chain at the position occupied by the proflavin molecule.g. sperms and Ova) are called gametic mutations. insert between two successive bases of a DNA strand. Classification of Mutations According to the Size and Quality According to size following two types of mutations have been recognized: Point mutation.BT0203 Genetics and Cytogenetics Gametic mutations. it is believed. The gametic mutations only form the base for the natural selection. hence.e.. The point mutations which occur due to addition of one or more extra nucleotides to a gene or cistron are called insertion mutations. then this type of mutations are called “point mutations”. 2. The point mutations may occur due to following types of subnucleotide change in the DNA and RNA. a single nucleotide or nucleotide pair.

3. such as adenine can in its rare state forms a bond with cytosine (besides thymine). while the purine guanine (G) is linked to the pyrimidine. each DNA base may have some altered uncommon molecular configuration... Tautomerization In a DNA molecule. it cannot be linked to its normal partner. Department of Genetic Engineering . guanine) or a pyrimidine (e.. They may be of following: (i) Transition. When a purine (e. thymine) is substituted by another pyrimidine base. The transitional substitution mutations occur due to tuatomerization. . Besides the common molecular configurations. The substitution mutation when involves the substitution or replacement of a purine with a pyrimidines of vice versa then type of substitution Rex Arunraj Assistant Prof. However. adenine (A) is linked to the pyrimidine. Such uncommon forms of DNA bases are generated by single proton shifts and are called rare states of tautomers.g. is called substitution mutation.g. When a base occurs in its rare or tatuomeric state. Transversion. Such an altered code word (triplet codon) may designate a different amino acid and may result in the production of a protein with a single amino acid substitution. Similarly. The substitution mutations alter the phenotype of an organism variously are of great genetical significance. provided the cytosine is in its normal state. a tautomeric shift may occur in thymine changing it form the keto (C = 0) form to the rare enol (COH) form. cytosine (C) by three hydrogen bonds. ademine) base of a triplet codon of a cistron is substituted by another purine base (e. The substitution mutation affects only a particular triplet codon. cytosine) then such kind of substitution is called transition. the purine. A tautomeric shift is believed to occur when the amino (NH 2) form of adenine is changed to an imino (NH) form..BT0203 Genetics and Cytogenetics replaced by another nucleotide.g. thymine (T). (e. normally. by two hydrogen bonds. a purine.g.

they can be recognized only by amino acid substitution in proteins. Effects of physical Conditions on Nucleotide Sequence High temperature and low pH value are known to affect depurination or loss of purine bases. the gross mutations occur due to rearrangements of genes within the genome and may be of the following types: 1.BT0203 Genetics and Cytogenetics mutation is called transversion mutation. 3. When changes involving more than one nucleotide pair. following two kinds of mutations have been recognized. However. The movement of a gene within the same chromosome is called inversion. 2. Moreover. We have still poor information about the mechanism of induction. then such mutations are called gross mutations. they may cause different types of phenotypic effects over the organisms. The rearrangements of genes may occur within a gene. The removal of a purine from a strand of DNA leaves a gap at that. identification and charectirization of trnsversion mutations. Freese in 1959. The rearrangements of gene may occur in number of genes per chromosome. Classification of Mutation According to the Origin According to the mode of origin. it would be possible for any of the four bases to insert in the complementary strand would contain a transversion. 3. At the time of replication. (ii) Inversion. Rex Arunraj Assistant Prof. Multiple mutations or gross mutations. Department of Genetic Engineering . or entire gene. especially when the gene loci may take place due to following method: (i) Translocation: Movement of a gene may take place to a non-homologous chromosome and this is known as translocation. The existence of transversion mutation was first of all postulated by E. it is extremely difficult to recognize transversion mutations genetically. Due to movement of a gene locus new type of phenotypes may be created. Two mutations within the same functional gene can produce different effects depending on gene whether they occur in the cis or trans position. B.

When ionizing radiations pass through matter. temperature) and chemicals. They are also called “background mutation” and have been reported in many organisms such as. Besides naturally occurring spontaneous mutations. The ejected electrons move at high speed. etc. Radiations. protons and other fast moving particles.e. become attach to other atoms and convert the ion into negatively charged ions. neutrons.e. knock other electrons free from their respective atoms and when their energy is dissipated. Both types of radiations induce mutations by following methods: (i) Ionizing radiations as mutagens. man. Relatively little is known about the mechanism by which ionizing radiations cause mutation. The second type is non-ionizing radiations such as ultraviolet and visible light. protons) and a surrounding constellation of negatively charged electrons. Drosophila. Department of Genetic Engineering . The spontaneous mutations occur suddenly in the nature and their origin is unknown. negatively charged particles (electrons) leaves atoms which are no longer neutral but are positively charged. the mutations can be induced artificially in the living organisms by exposing them to abnormal environment such as radiation. The substances or agents which induce artificial mutations are called mutagens or mutagenic agents. microorganisms (bacteria and viruses). electrons.BT0203 Genetics and Cytogenetics (1) Spontaneous mutations. in their turn. familiar that matter composed of atoms and atoms. To achieve their stable configuration (i. mice. The charges of atomic particles remain so balanced that normal is electrically neutral. are made up of a positively charged atomic nucleus (with neutrons. As. Mutagenic agents. alpha and beta rays. bread molds. The mutagenic agents are of the following kinds: A. The radiations which are important in mutagenesis are of two categories: one type is ionizing radiations such as X-rays and gamma rays. neutral charge) ions undergo many chemical Rex Arunraj Assistant Prof. certain physical conditions (i... they dissipate their energy in part through the ejection of electrons from the outer shell of atoms and the loss of these balancing. maize. (2) Induced mutations. Oenothera. The positively-charged atom is called ion.

(ii) Non-ionized radiations as mutagens: The ultraviolet (UV) light is a non-ionizing radiation which may cause mutation. one being the formation of chemical bonds between two adjacent pyrimidines molecules in a polynucleotide and particularly. ionizing radiations cause breaks in poly-sugar phosphate backbone of DNA and. It is reported that the rate of mutation is increased due to increase in temperature. their position in the DNA helix becomes so displaced that they can no longer from hydrogen bonds with the opposing purines and thus regularity of the helix becomes resorted. B. Dimerization: The ultraviolet radiation produces several effects on DNA. a state called excitation. The most effective wave length of ultraviolet for inducing mutations is about 2. Further. The rate of all chemical reactions is influenced by temperature. some of their electrons are raised to higher energy levels. Thus. inversion and translocation. Rex Arunraj Assistant Prof.Temperature as mutagen. between adjacent thymine residues. As the two thymine residues associate. oxygen is important in the formation of H 2O2 and HO2 in irradiated water and these products may induce breaks in DNA molecule. This will produce a T-A to C-G transition. Because. thus. The excited molecule becomes reactive and mutated and is called photoproducts. may result in thymine’s pairing with guanine. dimerization interferes with the proper base pairing of thymine with adenine. When a substance absorbs sufficient energy from the ultraviolet light. Temperature probably affects both thermal stability of DNA and the rate of reaction of other substances with DNA. For example. or dimerize to form a dimer. Department of Genetic Engineering . During breakage of DNA molecule due to ionizing radiation the active role of oxygen is predicted. causing chromosomal mutations such as break. addition. This is a wave length that is best absorbed by DNA and a wave length at which proteins absorb little energy. an increase of 10o C temperature increases the mutation rate two or three fold.BT0203 Genetics and Cytogenetics reactions and during these chemical reactions ionizing radiation is thought to cause mutation. deletion.600 Ao.

nitrous acid converts adenine into hypoxanthine and cytosine to uracil by deamination. called base analogues (e. methyl and ethyl methanesulphonate (MMS and EMS) and nitrosoguadine (NG) also have direct mutagenic effect on the DNA molecule. Any chemical substance that affects the chemical environment of chromosomes is likely to influence. the stability of DNA and its ability to replicate without error.g. The ability of chemicals to induce mutation was first of all demonstrated by Auerbach and Robson in 1947 using mustard gas and related compounds as the nitrogen and sulphur mustards. interfere with the integrity of the chromosome structure.)Closely resemble with certain DNA bases and are. tea and soft drinks). 2-aminopurine. Like the nitrous acid. dimethyl and diethyl sulphonate. It can affect the chromosomal DNA by following two ways: (1) Direct gene change. Effect of Chemical Mutagens on Nucleotide Sequence (a) Alteration in Resting Nucleic Acid Deamination: Some chemical substances such as nitrous acid causes transitional mutation due to oxidative deamination of DNA base. at least indirectly. Chemical mutagens. Department of Genetic Engineering . (2) Copy error. nitrogen mustard. thus. as they are the compounds which bind calcium and. phenol and carcinogens. epoxides.BT0203 Genetics and Cytogenetics C.. etc. Certain chemical compounds. Many chemical substances have been responsible to increase the mutability of genes. chloride is mutagenic for many organisms. caffeine (in coffee. Since then many chemical compound which are ordinarily considered to be non-toxic have been found to be mutagenic in certain specific situations. Certain chemical mutagens affect DNA directly. For example. 5bromouracil. where the amino group is replaced Rex Arunraj Assistant Prof. During analogues such as urethane triazine. therefore. They affect the constituents of DNA only when DNA is not replicating. formaldehyde. mustard oil and chloracetone in experiments with male Drosophilae melanogaster. A chemical mutation can cause mutation only when it enters in the nucleus of the cell. act as mutagens.

adenine is deaminated into hypoxanthine by nitrous acid. Base analogues. There is some in vitro evidence to indicate the BU immediately adjacent to an adenine in one of DNA strands causes the latter to pair with guanine. an A-T pair becomes and remains A-BU. These mutagens produce mutations in the following ways: (1) They add ethyl methyl groups to guanine. Department of Genetic Engineering . methyl methane sulphonate (MMS). For example. (3) The gap may also produce a deletion. 5BU behaves similar to the tautomer of thymine and pairs with guanine. This makes guanine the base analogue to adenine. such analogues may be incorporated into a replicating DNA stand. The loss of the base produces gaps in the DNA chain which may be filled with a wrong base. causing mutation. deamination converts cytosine to uracil. ethylethane sulphonate (EMS). Example of some most extensively studied alkylating agents include diethylsulphate (DES). which has pairing properties similar to thymine and in such a case G: C pair would be changed into A: T pair. This is known as depurination. But. Alkylating agents: Some alkylating agents carry one. 5-bromouracil (5BU) or its nucleoside 5bromodeoxyuridine (5-BUdR) in its usual (keto) form is a structural analogue of thymine (5-methyl uracil) and it will substitute for thymine. thus. Thus. (2) They remove the alkylated guanine. Similarly. 2-Aminopurine (2-AP) is another base analogue which is relatively undifferentiated purine that apparently can pair with cytosine and thymine. it may be incorporated opposite thymine during Rex Arunraj Assistant Prof. It is thought that 2-AP acts by “switching” pyrimidines: for example. producing mutation. in its rare (enol) state. Thus. (b) Alteration during Replication of Nucleic Acid 1. two or more alkyl groups in a reactive form and act as strong mutagens.BT0203 Genetics and Cytogenetics by hydroxyl (OH) group by the chemical mutagen. Certain chemical substances have molecular structure similar to the usual DNA bases that. dimethyl sulphate (DMS). This converts A: T to G: C. if they are available.

2. When a mutation occurs at a different site from the site where already primary mutation occurred and that mutated gene reverse the effects of primarily mutated gene. Inhibition of precursors of nucleic acid: There are some mutagents which interfere with the synthesis of nitrogen bases of nucleic acids such as purines or pyrimidines. Most mutations are forward type. The forward mutations are often corrected by error correcting mechanism. Often lack of one base either causes breaks or causes pairing mistakes. Department of Genetic Engineering . due to forward mutation the adenine is changed into guanine and backward mutation change guanine into adenine: Forward Adenine  reverse Guanine  Adenine (ii) Mutation suppressor. azaserine (a potent alkylating agent) is an inhibitor of pyrimidine synthesis. urethane induced chromosome breaks are inhibited by thymine 4. They may be of the following types: (i) Single site mutation. For example. In an organism when mutations create change from wild type to abnormal phenotype. Classification of Mutation According to the Direction According to their mode of direction following types of mutations have been recognized: (A) Forward mutations. so that an abnormal phenotype changes into wild type phenotype. They may be of following types: Rex Arunraj Assistant Prof. Some reverse mutations change only one nucleotide in the gene and are called single site mutations. (B) Reverse or back mutations. then such (secondary) mutations are called mutation suppressors. then that type of mutations are known as forward mutations. However. For example.BT0203 Genetics and Cytogenetics one round of replication and then pair with a cytosine at the next round to produce an AT GC transition.

coli. (i. (b) Intergenic suppressor. The extragenic suppressor mutation occurs in a different gene from that of the mutant gene. using the remaining opposite strand as a template. In photoreactivation type reverse mutation reversal of ultraviolet induced thymine dimers takes place by specific enzymes in the presence of visible light waves. the reverse mutation may also occur in the absence of light. (d) Excision repair or Dark reactivation. (c) Photoreactivation.BT0203 Genetics and Cytogenetics (a) Extragenic suppressor. During ultraviolet radiation a particular enzyme is selectively bound to the bacterial DNA. In an ultraviolet (UV) induced mutations. a gene called rec A (rec for recombination) is known which is necessary for recombination and is found to repair ultraviolet-induced thymine dimmers of a gene by a process called post replication recombinational repair . Classification of mutation According to Magnitude of Phenotypic Effect According to their phenotypic effects following kinds of mutations may occur: 1. Department of Genetic Engineering . in man the mutation disease aniridia (absence of iris of eyes) occurs due to dominant mutant gene. DNA ligase). Rex Arunraj Assistant Prof. on either side of the dimer which may be formed due to ultraviolet radiation and excises a short. For example. During photoreactivation the enzyme is activated by visible light and that cleaves the pyrimidine or purine dimers into monomers and restores their original forms. The intergenic suppessor mutation occurs in a different nucleotide within the same gene and shifts the reading frame back into register. (iii) DNA polymerase resynthesizes the missing segment. and (iv) the final gap is closed by some enzymatic rejoining process. In E. (ii) Another enzyme. Dominant mutations. The mutations which have dominant phenotypic expression are called dominant mutations.. possibly exonuclease widens the gap produced by the action of the endonuclease.e. 5. single strand segment of the DNA. According to Howard Flanders and Boyce (1964) dark reactivation includes following stages: (i) An enzyme possibly endonuclease makes a cut in the polynucleotide strand.

3. UAA or UGA). Most types of mutations are recessive in nature and so they are not expressed phenotypically immediately. Isoalleles. Department of Genetic Engineering . Classification of Mutation According to Consequent Change in Amino Acid Sequence. 4. If the substitution produces a protein that is active at one temperature (typically 30 o C) and inactive at a higher temperature (usually 40. Recessive mutations. 2. Lethal mutations result in the death of the cells or organisms in which they occur. 6. Supervital mutations. Rex Arunraj Assistant Prof. cause the improvement of biological fitness under certain conditions. According to their effects on the phenotype mutations may be classified as lethals. Missence mutations. 1. The phenotypic effects of mutations of a recessive gene are seen only after one or more generations.42o C). Nonsense or chain termination mutations. subvitals and supervitals. resulting in the production of a shorter protein. when the mutant gene is able to recombine with another similar recessive gene. Subvital mutations reduce the chances of survival of the organism in which they occur.BT0203 Genetics and Cytogenetics 2. 3. Lethal mutations. They produce identical phenotypes in homozygous or heterozygous combinations. Some mutations alter the phenotype of an organism so slightly that they can be detected only by special techniques. They arise when a codon for an amino acid is mutated into a termination codon (UAG. Mutant genes that give slightly modified phenotypes are called isoalleles. Temperature sensitive mutations or Ts mutations. They change the meaning of a codon. changing one amino acid into another. in contrast.

2. They change nucleotide but not the amino acid sequence because they affect the third position of the codon. Sex chromosomal mutations. 7. Mutation rate The frequency with which genes mutate spontaneously is called mutation rate. one gamete in 100. Mutations occur much more frequently in certain regions of the gene than in others. Classification of Mutation According to the types of Chromosomes. Autosomal mutations. According to the types of chromosomes. This is a silent mutation because it leaves the protein sequence unchanged. Silent mutations. Rex Arunraj Assistant Prof. Department of Genetic Engineering . they are the most versatile and useful mutations.000 to one gamete in million would contain a mutation at a given locus. 4. they are called conditional mutations. The favoured regions are called hot spots. The great majority of genes have mutation rate of 1X10-5 to 1 X 10-5. This type of mutation occurs in autosomal chromosomes. This type of mutation occurs in sex chromosomes. the mutations may be of following two kinds: 1. temperature-sensitive and chain termination mutations exhibit the mutant phenotype only under certain conditions. which is usually less important in coding. Most genes are relatively stable and mutation is a rate.BT0203 Genetics and Cytogenetics Since. viz.

This gave birth to a hybrid science. or gain. with genetic phenomena.e. animal husbandry and medicine. of a part of the chromosome set (aneuploidy) Loss. Chromosome mutations are inherited once they occur and are of the following types: A. Structural changes in chromosomes: 1. Department of Genetic Engineering . of whole chromosome set (euploidy) (a) (b) Loss of an entire set of chromosomes (haploidy) Addition of one or more sets of chromosomes (polyploidy) Exchange of parts between chromosomes of different pairs: Loss: deletion Addition: Duplication Rotation of as group of genes 180° within one chromosome: Changes in gene arrangement: Both types of changes (structural and numerical) in chromosomes can be detected not only with a microscope (i. let us consider two important features of chromosome behaviour : (1) During prophase I of meiosis.BT0203 Genetics and Cytogenetics CHROMOSOME ABERRATIONS Changes in Structure of Chromosomes The changes in the genome involving chromosome parts. 2. A. (a) inversion (b) translocation. or whole chromosome sets are called chromosome aberrations or chromosome mutations. or gain. Changes in number of genes (a) (b) 2.. especially those of chromosomes. cytologically) but also by standard genetic analysis. Loss. whole chromosomes. Rex Arunraj Assistant Prof. Structural Changes in Chromosomes For better understanding of the abnormalities of chromosome structure. called Cytogenetics which attempts to correlate cellular events. Changes in number of chromosomes: 1. Chromosome mutations have proved to be of great significance in applied biologyagriculture (including horticulture). B.

bears one block of genes in duplicate). an intermediate section or portion of chromosome is lost and it is caused by two Rex Arunraj Assistant Prof. Such loss of a portion of a chromosome (and of some genes) is called deletion. e. duplication involves addition of a part of chromosome (i. duplication homozygote. Types of Structural Changes in Chromosome Structural changes in chromosome may be of following types: 1. thus. Department of Genetic Engineering . showing strong tendency to join with broken ends. In terminal deletion a terminal section of a chromosome is absent and it is resulted by only one break.g. and 4. the broken chromosome ends are highly “reactive” or “stickly”. it is called structural heterozygote.. inversion in which broken segment reattached to original chromosome in reverse order. If gametes arise from the cells having a deleted chromosome. The chromosomes with deletions can never revert to a normal condition. Further. When both homologous chromosomes are involved. this deletion is transmitted to the next generation. deficiency or deletion which involves loss of a broken part of a chromosome. (2) structural changes usually involve chromosome breakage. While in the intercalary deletion. broken segment becomes attached to a homolog which.e. 2. Further. 1. Portions of chromosomes without a centromore (called acentric fragments) lag in anaphase movement and are lost from reorganizing nuclei or digested by nucleases. When only one homologous chromosome is involved. This property results in many curious structures observed in cells containing one normal chromosome set plus an aberrant set. deletion homozygote. 3.BT0203 Genetics and Cytogenetics homologous regions of chromosomes show a great affinity for pairing and they often go through considerable contortions in order to pair. a deletion can be terminal or intercalary (insterstitial). Deletion (or Deficiency) The simplest result of breakage is the loss of a chromosome.. translocation in which the broken segment becomes attached to a non-homologous chromosome resulting in new linkage relations. structural abnormalities can occur in both homologous chromosomes of a pair in only one of them. etc. these are called structural homozygotes.

i. this phenomenon is called pseudodominance. if a homozygous deletion is made.. INTERCALARY DELETION Deletion of some chromosome regions produce their own unique phenotypes. this is a small deletion and acts as a recessive lethal in this regard. However. smaller deletion in heterozygous condition can be tolerated by the organisms. Even individuals heterozygous for deletion (deletion in one of the homologous chromosomes) may not survive.BT0203 Genetics and Cytogenetics breaks – one on either end of the deleted region. the chromosome is broken into three pieces. Rex Arunraj Assistant Prof. bacteriophage and other organisms. Such cytological maps are often used to verify the genetic maps (based on linkage analysis) of these organisms. the region of deletion can be detected by the failure of the corresponding segment on the normal chromosome to pair properly. The cytological studies of pairing between normal and deleted chromosomes have helped a lot in finding out the relative position of genes in chromosomes. in the latter case. Further. it is lethal. Thus. in the presence of a deletion. In general. the middle one of which is lost and the remaining two pieces get joined again. If meiotic chromosomes in such heterozygotes are examined. a recessive allele of the normal homologous chromosome will behave like a dominant allele. a b c d e f a b c d e f c d e f g h g h g h g h g h a b c d e f a b e f TERMINAL DELETION Genetical effects of deletion. A good example of this is a dominant notch wing mutation in Drosophila. In fact. Department of Genetic Engineering . maize. it will be phenotypically expressed.e. so a “deletion-loop” results. The phenomenon of pseudodominance exhibited by deficiency heterozygotes has been utilized for the location of genes on specific chromosomes and in preparing cytological maps in Drosophila.

have malformation in the larynx. Tandem duplication. For example. In this case the duplicated regions are situated just by the side of the normal corresponding section of the chromosome and the sequences of genes are the same in normal and duplicated region. if the sequence of genes in a chromosome is ABC. at meiosis the chromosome bearing the duplicated segment forms a loop to maximize the juxtaposition (during pairing) of homologous regions. Department of Genetic Engineering . the sequence of genes due to reverse tandem duplication will be ABC. the sequence of genes in tandem duplication will be ABC. moon faces. Displaced duplication. They are also mentally retarded (IQ below 20). the French name “cridu chat” (cry of the cat) syndrome (first described by Lejeune et al. Reverse tandem duplication.BT0203 Genetics and Cytogenetics Examples of pseudodominance (deletion) Human babies missing a portion of the short arm of chromosome 5 (autosome) have a distinctive cat-like cry. In this case the duplicated region is not situated adjacent to the normal section. 1963). DEF DEFGH. Depending on whether the duplicated portion is on the same side of the Rex Arunraj Assistant Prof. saddle noses. Duplication The presence of a part of a chromosome in excess of the normal complement is known as duplication. the sequence of genes in the duplicated region of a chromosome is just the reverse of a normal sequence. hence. Here. DEFGH (The full stop depicts the Centro mere) and if the chromosomal segment containing the genes DEF is duplicated. Extra segments in a chromosome may arise in a varsity of ways such as follows: 1. 2. If duplication is present only on one of two homologous chromosomes. malformed low-set ears and microcephally (small head). In the above mentioned example. small mandibles (micrognathia). therefore. DEF FEDGF.. 3. Thus. due to duplication some genes are present in a cell in more than two does. 2.

Here. The net result of inversion is neither a gain nor a loss in the genetic material but simply a rearrangement of Rex Arunraj Assistant Prof.RST.C. Inversion Inversion involves a rotation of a part of a chromosome or a set of genes by 180° on its own axis. if ABC. Extra-chromosomal duplication.DEFGH and LMNOPQ.RST DEF). and (3) another duplication causes hairy wing (Hw) Genetic redundancy.GH and LMN DEF. a transposed duplication will result into chromosomes with gene sequence ABC. 3.. DEFH Heterobranchial duplication = A. Due to duplication.g. there occur unequal crossing over which results in deletion and reduplication which produce distinct phenotypes as shown by the following examples : Duplications of Drosophila lead to following phenotypic effects : (1) a reverse repeat in chromosome 4 causes eyeless dominant (Ey). 5.RST represent the gene sequences of two nonhomologous chromosomes.e. DEFOPQ. may protect the organism from the effects of a deleterious recessive gene or from an otherwise lethal deletion. Department of Genetic Engineering .OPQ. Transposed duplication.RST) or terminal (i.DEFGH 4. the displaced duplication can be termed either homobranchial or heterobranchial. Such a transposed duplication may be either interstitial (e. the duplicated portion of chromosome becomes attached to a non-homologous chromosome.DEFG. of which duplication is one type.BT0203 Genetics and Cytogenetics centromere as the original section or on the other side.DEFB.. Genetical effects of duplication. (2) a tandem duplication in chromosome 3 causes confluens (Co) resulting in thickened veins. LMN OPQ. For example. It essentially involves occurrence of breakage and reunion. LMN. Homobranchial duplication =ABC. In the presence of centromere the duplicated part of a chromosome act independent chromosome. Example.

This results in an inverted chromosome having segments 1-2-5-4-3-6. Fertility of inversion homozygotes and sterility of inversion heterozygotes lead to establishment of two group (or varieties) which are mutually fertile but do not breed well with the rest of the species. only a rearrangement (i. the broken segment of one chromosome gets inserted interstitially in a nonhomologous chromosome. They involve a single break in a chromosome. change in the sequence and position of a gene). Advantage of inversions. Department of Genetic Engineering . The broken piece gets attached to one end of a nonhomologous chromosome. Translocation The shifting or transfer of a part of a chromosome or a set of genes to a nonhomologous one. Homologous chromosomes..e. In this type of translocation. 2. The location of the inverted segment can be detected cytologically in the meiotic nuclei of such heterozygotes by the presence of an inversion loop in the paired homologs. Both varieties evolve in different directions and later become reproductively isolated species. An inversion can occur in the following way: suppose that the normal order of segments within a chromosome is 1-2-3-4-5-6.BT0203 Genetics and Cytogenetics the gene sequence. breaks occur in regions 2-3 and 5-6 and broken piece is reinserted in reverse order. If the centromere is not included in the inversion it is called paracentric inversion and when inversion includes the centromere it is called pericentric inversion. duplications and other curious configurations. An inversion heterozygote has one chromosome in the inverted order and its homologue in the normal order. 4. 1. Shift translocation. There is plenty of cytological evidence to prove that such evolutionary mechanisms have and are operating in Drosophila and a number of other orgasisms. There is no addition or loss of genes during translocations. Simple translocation. Translocations may be of following three types. However. with identical in meiosis. is called translocation. crossing over in inversion heterozygotes produce deletions. The location of the centromere relative to inverted segment determines the genetic behaviour of the chromosomes. Rex Arunraj Assistant Prof.

35 and 56. 8x). Except diploids. many commercial fruits and ornamental plants. Types of polyploidy Rex Arunraj Assistant Prof. pentaploid. Outcomes of reciprocal translocation. e. a large metacentric chromosome is shortened by one-half in length to an acrocentric one.BT0203 Genetics and Cytogenetics 3. In this case. a segment from one chromosome is exchanged with a segment from another nonhomologous one.Changes in number of chromosomes (ploidy) Polyploidy Any organism with more than more than two genomes (2x) is called a polyploidy. The exchange of chromosome parts between nonhomologous chromosomes creates new linkage relationships. tetraploid. Generally. For example the rose genus Rosa includes species with the somatic numbers 14. Many plant genera include species whose chromosome numbers constitute a euploid series. strawberries (octaploid. The translocation homozygotes may have normal meiosis and in fact. are difficult to detect cytologically unless morphologically dissimilar chromosomes are involved. 28. Two types of translocations have been recognized: homozygous and heterozygous. These numbers are the multiples of 7. triploid. wheat (hexaploid 6x). 21. this is a euploid series of the basic monoploid number 7.. For example. Thus. Ploidy levels higher than tetraploid are not commonly encountered in natural populations. rest of these belongs to polyploidy category. so that in reality two translocation chromosomes are simultaneously achieved. where as the small chromosome becomes a large one. Such translocations also drastically change the size of a chromosome as well as the position of its centromere. polyploidy is common in plants (more common in monocots) but rare in animals. Therefore. Reciprocal translocations. The translocation heterozygotes produce both translocated and normal chromosomes and exhibit characteristic cytological and genetical effects. hexaploid and octaploid species. Department of Genetic Engineering . B. which gives diploid. translocation heterozygotes are marked by considerable degree of meiotic irregularity. but our most important crops and ornamental flowers are polyploids. or banding patterns differ markedly.g.

which consist of same basic set of chromosomes multiplied. (ii) allopolyploids. radium and X-ray) and temperature shocks. during cell divisions. many important crop plants include autotetraploids such as rye (Secale cereale). For example. grapes. corn (Zea mays).Colchicum autmunale and C.g.BT0203 Genetics and Cytogenetics These are the three different kinds of polyploids: (i) autopolyploids. and an autotetraploid. When they are found in nature. if a diploid species has two similar sets of chromosomes or genomes (AA). Colchicine Colchicine is a drug (i. (iii) autoallopolyploids. red clover (Trifolium pretense). berseem (Trifolium alexandrium). snapdragons (Antirrhinum). Phlops. their autopolyploidy is deduced by their meiotic behaviour. These inducers usually disturb the mitotic spindle and cause nonsegregation of already duplicated chromosomes. will have four such genomes (AAAA). Induced autopolyploidy The autopolyploidy have been induced in many plant and animal cells by artificial means such as chemical (e. Some of common examples of autotriploid crop plants.. are seedless varities of watermelons.g. Similarly.luteum) and its aqueous solution is found to prevent the formation and organization of spindle fibres. an autotriploid will have three similar genomes (AAA). marigolds (Tagetes). Oenothera lamarckiana (which was recognized as mutation by Hugo de vries). e.). mercury chloride. polyploids. colchicines. etc. Department of Genetic Engineering . hexachlocyclohexane. (i) Origin and production of autopolyploids The autopolyploids may occur in nature or may be produced artificially. apples. tomato. radioactive substances. sugar beet.e. sulphanil amide. grapes and banana. (a) Autopolyploids The autopolyploids are those. which are mainly produced by artificial methods. chloral hydrate.. an alkaloid obtained from the corms of plants. so the metaphase chromosomes of the affected cells (called C-metaphase or colchicine metaphase) do not move to a metaphase plate and remain Rex Arunraj Assistant Prof..

the water content increases which leads to a decrease in osmotic pressure. (2) Due to lower rate of cell division. (4) At higher ploidy level. the size of lower epidermis of leaf of a tetraploid Saxifraga pencylvanica was found greater than the diploids. leaves. These triploids are obtained from seeds by colchicines treatment. Even the process of cytokinesis is prevented by colchicines and with duplications of chromosomes the number goes on increasing. Hitoshi Kihara. then it is called allopolyploidy and the resultant species is called an allopolyploid. (ii) Effects of autopolyploidy Autopolyploidy results in gigantism of plant cells. This results into loss of resistance against frost. the time of blooming of an autopolyploid is delayed. Dr. Polyploid varieties with an even number of genomes (e. the fertility level and seed set are low. Let A represent a set of chromosomes (genome) in species X.g.. This leads to a decrease in auxin supply and a decrease in respiration. the adverse effects become highly pronounced and lead to the death of the plants. (3) Due to slow growth rate. triploids) are highly sterline. flowers and fruits of an polyploid are larger in size than a diploid plant.g. Some of the significant effects of autopolyploidy are as follows : (1) With the increase of cell size. its effects are limited to divided and meristematic cells.e. such as autooctoploids. Department of Genetic Engineering . As colchicine interferes with spindle formation. i. so seedless fruits can be produced by using triploids as in case of seedless watermelons which were produced by using triploids as in case of seedless watermelon which were produced by a Japanese scientist.BT0203 Genetics and Cytogenetics scattered in the cytoplasm.. and let B represent another genome in a species Y. the plants growth rate in decreases. Uses of induced polyploidy Since in the induced polyploids. The F1 hybrids of these species then would have one A genome and another B genome. For example. The Rex Arunraj Assistant Prof. Allopolyploids When the polyploidy results due the doubling of chromosome number in a F 1 hybrid which is derived from two distinctly different species. etc. tetraploids) are often fully fertile whereas those with an odd number (e..

Sears and also by H. Common hexaploid wheat and tetraploid cotton furnish two such examples.BT0203 Genetics and Cytogenetics doubling of chromosomes in the F1 hybrids will give rise to allotetraploids with two A and B genomes.spelta. a Russian geneticist. Triticale is an artificial allopolyploid which has been derived by crossing wheat (Triticum) and rye (Secale). When the synthesized hexaploid wheat was crossed with naturally occurring T. Triticale Triticale (Triticosecale Wittmack) is the first man made cereal which has been developed in recent years and is cultivated on about one million hectares of land throughout the Globe for the commercial use. Among these sterile F1 hybrids. Synthesized Allopolyploids To find out the origin of naturally occurring allopolyploids some cytogeneticists produced certain allopolyploids in laboratory by employing artificial means. Depending upon whether Triticum is a tetraploid (2n = 4x =28) or hexaploid (2n = 4x = 42). Triticum dicoccoides. Aegilops squarrosa (diploid .S. In 1927.spelta. This suggested that hexaploid wheat must have originated in the past due to natural hybridization between tetraploid wheat and goat grass followed by subsequent chromosome doubling. 2n = 18) and cabbage (Brassica oleracea. G. Raphanobrassica is a classical example of allopolyploidy or amphipolyploidy. These fertile tetraploids were called Raphanobrassica. Department of Genetic Engineering .McFadden and E.R.Kihara. Triticum spelta Triticum spelta is a hexaploid wheat which was artificially synthesized in 1946 by E. (tetraploid : 2n = 28) with goat grass. They crossed an emmer wheat. 2n = 18) and in F 1 got sterile (diploid) hybrids. he found certain fertile plants which were found to contain 36 chromosomes. This artificially synthesized hexaploid wheat was found to be similar to the primitive wheat T.D. the F1 hybrid was completely fertile.Karpechenko performed a cross between radish (Raphanus sativum. 2n = 14) and doubled the chromosome number in the F 1 hybrid. one would get hexaploid Rex Arunraj Assistant Prof.

etc. Likewise corn meal of a tetraploid maize seed contains 40 per cent more than vitamin A than cornmeal from a diploid plant. respectively. The polyploidy is invariably related with gigantism. such allopolyploids are called segmental allopolyploids . xylem. Therefore. Department of Genetic Engineering . stomata. (iv) Evolution through polyploidy. cells. Segmental allopolyploids Different genomes of some allopolyploids are not quite different from each other. leaves. Rex Arunraj Assistant Prof. only diploid rye (2n = 4x = 14) was used. Interspecific hybridization combined with polyploidy offers a mechanism whereby new species may arise suddenly in natural populations. This indicates that segments of chromosomes and not the whole chromosomes are homologous.BT0203 Genetics and Cytogenetics triticale (2n = 6x = 42) or octaploid triticale (2n = 8x = 56). Phenotypic Effects of Polyploidy The increase in the genome’s size beyond diploid level is often caused following detectable phenotypic characteristics in a polyploidy organism: (i) Morphological effect of polyploidy. In each case. The most important effect of polyploidy is that it reduces the fertility of polyploid plants in variable degrees. (ii) Physiological effect of polyploidy. The ascorbic acid content has been reported to be higher in tetraploid cabbages and tomatoes than in corresponding diploids. Consequently in these polyploids chromosomes belonging to different genomes do pair together to some extent. (iii) Effect on fertility of polyploidy.The segmental allopolyploids are intermediate between autopolyploids and allopolyploids and can be identified by their peculiar meiotic behaviour. The polyploidy plants have been found to contain large-sized pollen grains. The polyploid plants are more vigorous than diploids.

chloroplasts have a characteristic pattern of inheritance which does not resembles genes of nuclear chromosomes and are hence called as non-Mendelian. regardless of the genotype and phenotype of pollen parent and likewise. Mirabilis Jalapa. Department of Genetic Engineering . extra chromosomal. Correns further reported that flowers from the variegated branches yielded mixed progeny of green. The plants from the white or pale seedings die because the lack chlorophyll and cannot carry photosynthesis. Correns in 1908 in the four o’ clock plant. white (pale) and variegated plants in widely varying ratios. cytoplasmic and extra nuclear inheritance. In contrast to other higher plants. non-chromosomal. mitochondria. Chloroplasts and mitochondria are organelles that contain their own DNA and protein synthesizing apparatus. so the white or pale parts of plant survive by receiving nourishments from green parts. A widely held theory concerning their origin proposes that they were once infectious endosymbiotic prokaryotes that involved such as dependence on the gene products of the host that they are no longer able to function autonomously. Because the chlorophyll pigment of chloroplast is related with photosynthesis of food and leucoplasts are incapable to perform photosynthesis. (a) Chloroplast inheritance in variegated four o’ clock plant. flowers from the white or pale branches produced only white or pale seedings regardless of genotype and phenotype of pollen parent.BT0203 Genetics and Cytogenetics Extra-nuclear inheritance by cellular Organelles Cytoplasmic extra nuclear genes or DNA molecules of plastids. (2) White (pale) leaves and branches having no chloroplast. Correns reported that flowers in green branches produced only green offspring’s. The cytoplasmic or extra nuclear inheritance of colour in plant by plasticides was first of all described by C.. A study of the egg during oogenesis in Mirabilis reveals that the ooplasm contains plastids like cytoplasm of other Rex Arunraj Assistant Prof. (3) Variegated branches having leucoplast in white (pale) areas and chloroplast in green patches. Miabilis contains three types of leaves and parts: (1) Full green leaves or branches having chloroplast. The irregularly of transmission from variegated branches could be understood by considering cytoplasmic genes (plasmagenes) of plastids.

the F1 progeny would always be male sterile. Mitotic segregation. its ooplasm will contain coloured plastids. reveals that pollen contains very little cytoplasm which in most cases is devoid of plastids. if the female parent is male sterile (having plasmagene for male sterility). Variegated branches of Mirabilis Jalapa produce three types of eggs: some contain only white chloroplasts. having homozygous dominant restorer genes. however. Without the plastids. though the male sterility is fully controlled by the cytoplasm. if the male Rex Arunraj Assistant Prof. its ooplasm will contain white plastids. In maize and many other plants. white plastids only or a mixture of coloured and white plastids. so it is called cytoplasmic segregation and recombination (its acronym is CSAR). pollen cannot affect this aspect of the offspring’s phenotype. Their F1 progeny would be male fertile Rr. but a restorer gene if present in the female parent in the nucleus. In mitotic segregation since both segregation and recombination of organelle genotype takes place.BT0203 Genetics and Cytogenetics plant cells. some form of cytoplasmic segregation occurs that segregate the chloroplast types into pure cell lines. thus. some contain only green chloroplasts and some contain both types of chloroplasts. (b) Cytoplasmic male sterlity (CMS). However. if derived from variegated tissues. if the female parent is male sterile (due to plasmagene of male sterility) then the nuclear genotype of the male parent will determine the phenotype of F1 progeny. For example. A study of the pollenogenesis. cytoplasmic control of male sterility is known. In the subsequent mitotic divisions. (c) Cytoplasmic genetic male sterility. In certain plants. will restore fertility. because the cytoplasm is mainly derived from the egg which is obtained from the male sterile female parent. This process of sorting might be described as “mitotic segregation” of this is a pure extra nuclear phenomenon. its cytoplasm may contain coloured plastids only. Department of Genetic Engineering . producing the variegated phenotype in the progency individual. if male sterile female parent contains recessive nuclear genotype rr of restorer gene and male parent is RR. If the egg cell is derived from green plant tissues. if derived from white plant tissues. In such cases. Thus.

C and S).. Later on.BT0203 Genetics and Cytogenetics parent is male fertile rr. in maize the following four types of cytoplasms have been recognized: the normal (N) cytoplasm and three types of male sterile cytoplasms (T. especially on large scale. The recent studies of mitochondria in these cytoplasm revealed that the factors responsible for cytoplasmic male sterility are located in mitochondrial DNA (mt DNA) and mt DNA of N. the F 1 progeny would be male sterile rr. Male sterile lines can bear seeds only after crosspollination. Department of Genetic Engineering . The cytoplasmic male sterility (CMS) of C and S type can be reversed by nuclear storer genes. T. For this reason they are useful in raising hybrid seeds. in maize expression of male sterility depends on an interaction between nuclear and extra chromosomal genes. C and S cytoplasms are found to be different. however. If the F 1 male fertile heterozygote (Rr) is test crossed with male fertile progeny with 50 per cent male fertile and 50 per cent male sterile will be obtained. Male sterile F1 (O) Intercross or selfing Male fertile Male fertile Male fertile (O) Male fertile Rex Arunraj Assistant Prof. Since. Parents . the CMS-T cannot.

Department of Genetic Engineering .hybrid Male sterile -pure Inheritance pattern of genetic male sterility Rex Arunraj Assistant Prof.BT0203 Genetics and Cytogenetics F2 25% Male fertile -Pure 50% 25% Male fertile.

is undoubtedly important in the evolution of bacteria just as it is in the evolution of eukaryotes. The uptake of DNA molecules by recipient bacteria is an active. (1) Transformation involves the uptake of naked DNA molecules from one molecule from one bacterium (the donor cell) by another bacterium (the recipient cell). and Haemophilus influenzae. D. (3) Conjugation is the process during which DNA from a donor or male cell is transferred to a recipient or female cell through a specialized sex plus or “conjugation tube”. making possible the subsequent recombination events. It does not involve passive entry of DNA molecules through permeable cell walls and membranes (although this type of uptake of DNA molecules may be induced by experimental manipulations of bacteria in the laboratory. (2) Transduction occurs when bacterial genes are carried from a donor cell to a recipient cell by a bacteriophoge. and McCarty’s (1944) proof that the “transforming principle” (the cellular component mediating transformation) is DNA. Department of Genetic Engineering .BT0203 Genetics and Cytogenetics UNIT – V GENETIC TRANSFER Recombination. TRANSFORMATION Transformation was discovered in pathogenic strains of Diplococcus pneumoniae by Griffith in 1928. such as in the Escherichia coli recombinant DNA cloning experiments). transformation does not occur “naturally” in all species of bacteria. The details of Avery. The most obvious difference between these three processes is the most of transfer of DNA from one cell to another. Macleod. Even in the species. pneumoniae. energy-requiring process. Most of the studies on transformation have been done with three species. Three different processes have evolved that mediate transfer of genetic material from one bacterium to another. only in those species possessing the enzymatic machinery involved in the active uptake and recombination processes. Thus. Only Competent cells. all cells in the given population are not capable of active uptake of DNA. which possess a so-called “competence factor” (probably a cell-surface protein or enzyme involved in binding or Rex Arunraj Assistant Prof. Bacillus subtilis. however.

BT0203 Genetics and Cytogenetics in taking up DNA).000-10.albeit homologous . Not all bacteria can become competent. apparently prior to the completion of cell wall synthesis. DNA is not exchanged between distantly related microbes. This recombination replaces the gene in the host with a variant . Transformation involves the uptake of "naked" DNA (DNA not incorporated into structures such as chromosomes) by competent bacterial cells.000 nucleotides. The single-stranded DNA may recombine with the host's chromosome once inside the cell. The proportion of bacteria in a culture that are in the physiologically competent state varies with growth conditions and the stage of the growth curve (becoming maximal in late log-phase). While transformation occurs in nature. typically containing calcium salts. the extent to which it contributes to genetic diversity is not known. are capable of serving as recipients in transformation. in general.gene. At the entry site. endonucleases cut the DNA into fragments of 7. (2) irreversible uptake of the donor DNA (at which time the donor DNA becomes resistant to DNase in Rex Arunraj Assistant Prof. The process of transformation can be divided into several stages: (1) reversible binding of doublestranded DNA molecules to receptor sites on the cell surface. DNA from a closely related genus may be acquired but. Department of Genetic Engineering . and the doublestranded DNA separates into single strands. Cells are only competent (capable of taking up DNA) at a certain stage of their life cycle. Genetic engineers are able to induce competency by putting cells in certain solutions.

In most transformation experiments. One attractive model. replacing a segment of one strand of the recipient chromosome). is specific for homologous DNA. Although very small fragments of DNA taken up by competent cells. proposes that a specific exonuclease (or DNA “translocase”) pulls one strand of donor DNA into the cell using energy derived from the degradation of the complementary strand. Steps (2) and (3) may well be coincident effects of a single process. The first three steps in transformation-binding. donor DNA fragments are about 20. Genetic mapping by transformation Rex Arunraj Assistant Prof. however. In fact. During integration. and (5) the segregation and phenotypic expression of the integrated donor gene or genes in the recombinant (“transformed”) cell. Whether degradation of the complementary strand or DNA actually occurs during uptake or immediately after uptake is uncertain. the previous degradation of one strand is at random) of donor DNA is physically inserted into the recipient chromosome. considerable evidence suggests that these processes may vary in different species.000 nucleotide-pairs (or about 1/200 of the total chromosome) in length. a minimum length of about 500 nucleotide-pairs appears to be required for integration to occur. This is not to say that the integration of segments of heterologous (foreign) DNA never occur. Moreover. Department of Genetic Engineering .BT0203 Genetics and Cytogenetics the medium). it does so at frequencies very much lower than the frequencies observed using homologous DNA. competent bacteria will carry out these three processes equally with other foreign DNAs. This means that mapping experiments can be done using transformation only if the genetic markers employed are located close together on the host chromosome. However. the integration. uptake. or DNA recombination step. (4) integration (covalent insertion) of all or part of the single strand of donor DNA into the chromosome of the recipient. a single strand (either strand. for which there is supporting evidence in the case of Pneumococcus. and degradation of one strand of the double stranded DNA – are not specific for homologous DNA. (3) conversion of the doublestranded donor DNA molecules to singlestranded molecules by nucleotic degradation of one strand.

The phage can then break open (lyse) the cell. In this case. two genes are closely linked. Thus. or sexduction experiments. Thus. Genetic markers can also be ordered be means of three-factor transformation experiments using the same rationale as in threefactor transduction. Since transformation of any single marker occurs with allow frequency. The frequency with which two genetic markers are cotransformed can thus be used as a crude estimate of the linkage distance between them. The probability of two such independent events occurring together will equal the product of the probability of each occurring alone. Early in the infective cycle the phage encodes an enzyme that degrades the DNA of the host cell. conjugation. Some of these fragments of bacterial DNA are packaged within the bacteriophage particles. TRANSDUCTION Principle Transduction is another method for transferring genes from one bacterium to another. using an a + b+ donor and an a b recipient) will require two independent transformation events (uptake and integration of one DNA molecule carrying a+ and another molecule carrying b+). A bacteriophage infection starts when the virus injects its DNA into a bacterial cell. transferring the bacterial genes. a phage that contains bacterial genes can continue to infect a new bacterial cell. they may be carried on a single molecule of transforming DNA. If. double transformants can be formed by the uptake and integration of one molecule of donor DNA carrying both genes. When released from the infected cell. taking the place of phage DNA. double transformants for the two genes (say a to a+ and b to b+. on the other hand. this time the transfer is mediated by bacteriophages (bacterial viruses. Sometimes genes transferred in this manner become integrated into the genome of their new bacterial Rex Arunraj Assistant Prof. double transformants may be formed at a frequency approaching the frequency of single transformants in comparable single-marker experiments. if two genes or genetic markers are very closely linked. also called phages). double independent transformation events of this type will be extremely rare. Bacteriophage DNA is replicated and then packaged within the phage particles. they will never be carried on the same molecule of transforming DNA. The bacteriophage DNA may then direct the synthesis of new viral components assembled in the bacterium.BT0203 Genetics and Cytogenetics If two genes are far apart on the chromosome. Department of Genetic Engineering .

Transduction occurs in a wide variety of bacteria and is a common mechanism of gene transfer.BT0203 Genetics and Cytogenetics host by homologous recombination. occurs when a bacteriophage particle carries a segment of the chromosome from one bacterium (the donor) to another bacterium (the recipient). Since all the genes of the donor are represented in a population of these transducing particles (although any one transducing phage contains only one segment of host DNA. called transducing particles. representing 1/100 to 1/50 of the total donor of the chromosome). Two very different types of transduction are known. this type of transduction was named “generalized” transduction. facilitating subsequent recombination of the genetic markers of the two cells. a recombination event involving the host chromosome and the phage chromosome occurs. Specialized transduction is so named because a given virus transduces only genetic markers of the host that are located Rex Arunraj Assistant Prof. Department of Genetic Engineering . Such transduced bacteria are not lysed because they do not contain adequate phage DNA for viral synthesis. (1) In generalized transduction. Specialized transducing particles thus always contain both phage and bacterial DNA. (2) In specialized transduction (also called restricted transduction). Generalized transducing phages can therefore transport any gene of the donor cell to the recipient cell. producing a phage chromosome containing a segment a bacterial DNA. Transduction. Lederberg in 1952. a random segment of bacterial DNA is “wrapped up” during phage maturation or along with phage chromosome in a few “progeny” particles.Zinder and J. discovered by N.

E.coli. Virulent phages always multiply and lyse the host cell after infection. Generalized transducting particles are produced during the lytic cycles of these phages. Thus. except that the integrated segment is double-stranded. Salmonella phage P22. or (2) remain free in the cytoplasm. GENERALIZED TRANSDUCTION Bacteriophages have been classified into two types on the basis of their interactions with the bacterial cell. usually mediates transduction of the gal and bio genes of E. After a transducing phage injects a fragment of DNA into a recipient cell. Temperate phages have a choice between two life-styles after infections. they are Rex Arunraj Assistant Prof. it will not replicate and will be transmitted to only one progeny cell during each cell division. for example. they can (2) enter the lysogenetic pathway during which their chromosomes are integrated into the chromosomes of the host and replicate like any other segments of the host chromosomes.coli phage P1.BT0203 Genetics and Cytogenetics in one small region of the bacterial chromosome. the probably of the cell being doubly transduced for markers carried in two different transducing particles in negligible. Of the generalized transducing phages. that DNA may either (1) be integrated into the host chromosome in a manner similar to the integration of transforming DNA. Gene mapping by transduction Transducing particles are produced at a low frequency. Cells carrying nonintegrated transducing fragments are called abortive transductants. If it is not integrated. Bacteriophage lambda. and Bacillus subtilis phages PBS1 and SP10 have been extensively used for genetic fine structure maping. Department of Genetic Engineering . even they are not integrated. The genes located on the transduced chromosome fragments may be expressed. Generalized transduction is mediated by some virulent bacteriophages whose chromosomes are not integrated at specified attachment sites on the host chromosome. Only one out of 10 5-107 of the “progeny” particles present in a lysate contains bacterial DNA. They can either (1) enter the lytic cycle. the best-known specialized transducing phage. or. during which they reproduce and lyse their host cells just like virulent phages. alternately. (If cells are simultaneously infected with 100 or more phage particles.

(2) markers b+ and c+ are cotransduced. because the Rex Arunraj Assistant Prof. If (1) markers a+ and b+ are cotransduced. As such. This site specific recombination event results in the covalent linear insertion of the page chromosome into the chromosome bacterium. those involved in viral reproduction and lysis of the host. involves a recombination event between the circular intercellular form of the phage chromosome and the circular bacterial chromosome at specific attachments sites on the two chromosomes. they are examples of genetic elements called episomes. The chromosomes of temperate phages of this type are thus capable of both (1) autonomous replication (replication independent of the replication of the host chromosome) and (2) integrated replication (replication as a segment of the host chromosome). genetic markers can be ordered by cotransduction patterns. A bacterium harboring a prophage is said to be lysogenic. More frequently. the prophage-host relationship is called lysogeny.) The cotransduction of two or more genetic markers therefore indicates that the markers are relatively closely linked.BT0203 Genetics and Cytogenetics rapidly killed by a process called “lysis-from-without”. This site specific recombination event results in the covalent linear insertion of the phage chromosome and the circular bacterium chromosome at specific attachment sites on the two chromosomes. but (3) markers a+ and c+ are not cotranduced. SPECIALIZED TRANSDUCTION Specialized transduction is mediated by temperate bacteriophages whose chromosomes are able to integrate at one or a few specified attachment sites on the host chromosome. the phage chromosomes are called a prophage. are repressed (turned off) when the chromosome is in the prophage state. Occasionally. Integration of the chromosome of a specialized transducing phage. then the order of the three markers must be a+-b+-c+. such as the coliphage lambda. A lysogenic cell is immune to secondary infections by the same virus (homologous to the prophage). In its integrated state. and the frequency of cotransduction of any two markers is indicative of the degree of linkage between them. however. Department of Genetic Engineering . (The mechanism by which the prophased genes is repressed). three-factor transduction experiments must be used to unambiguously order genetic markers. which apparently results from simply punching too many holes in the cell membrane. The lytic genes of the virus.

the prophage is excised from the host chromosome and commences replicating autonomously. The excision process is site specific. When this happens. Department of Genetic Engineering . however.BT0203 Genetics and Cytogenetics lytic genes of the infecting virus will be repressed just as those of the prophage are repressed. Such “mistakes” during prophage excision are responsible for the formation of specialized transducing particles. Thus specialized transduction is restricted to the transfer of genes located within a short distance on each side of the prophage attachment site. for example. The excision process is usually very precise in cutting out the phage chromosome in exactly the form in which it existed prior to its integration. by irradiation with ultraviolet light. a portion of the phage chromosome is left in the host chromosome and a portion of the bacterial chromosome is excised with the phage DNA. Only host genes located close to the site of prophage insertion can be excised with the phage DNA and packaged in “phage” particles. the excision event occurs at a site other than the original attachment site. During the switch from the lysogenic state to lytic growth. Phage lambda integrates between the gal genes (required for the utilization of Rex Arunraj Assistant Prof. Occasionally. Such transitions can be also induced. like the integration process. The site-specific integration and excision processes are catalyzed by enzymes encoded by phase genes. Temperate phages undergo rare (about one in105 cell divisions) spontaneous transitions from the lysogenic or prophage state to the lytic state.

just as in transformation and transduction. called F . lambda thus usually only transduces gal and bio markers.coli trp genes (required for the synthesis of the amino acid trytophan) and transduces trp markers.L. no transducing particles are present in the phage lysates. The synthesis of these F pili is controlled by several (nine. based on current data) genes that are carried by a small circular molecule of DNA or “minichromosome” (about 94. in which it replicates independently of the chromosome. DNA is transferred from a donor cell to a recipient cell through a specialized intercellular connection.coli chromosome. The transfer of genetic information is thus a one-way transfer during conjugation. on the other hand. CONJUGATION Conjugation was discovered in 1946 by J.cells (recipient cells). that forms between them. Lederberg and E. or conjugation tube. Specialized tansducing phage O 80. Tatum (1958 Nobel Prize co recipients). rather than a reciprocal exchange of genetic material. During conjugation. integrates near the E. The frequency of transducing particles in lysates produced by induction of induction of lysogenic cells is about one in 106 progency particles. The F factor can exist in two different states: (1) the autonomous state. If bacteria are infected by specialized transducing phages under conditions where only lytic infections occur. Cells that have the capacity to serve as donors during conjugation are differentiated by the presence of specialized cell-surface appendages called F pili.BT0203 Genetics and Cytogenetics galactose as an energy source) and the bio genes (essential for the synthesis of biotin) on the E. The donor and recipient cells are sometimes referred to as male and female cells. in which it is Rex Arunraj Assistant Prof. and (2) the integrated state.500 nucleotide-pairs long) called on F factor (for fertility factor. also called “sex factor” and “F plasmid”). This is indeed the case. Department of Genetic Engineering . If specialized transducing particles are formed during prophage excision. only phage lysates produced by induction of lysogenic cells should have transducing activity. respectively). Cells carrying an F factor (donor cells) form conjugation tubes and initiate DNA transfer making contact with cells not carrying an F factor.

cells results in virtually all the cells in the new population becoming F+.recipient cell. In the integrated state. A donor cell containing the F factor in the autonomous state is called an F+ cell. Rex Arunraj Assistant Prof. only the autonomous F factor is transferred. Both exconjugants (cells that have been involved in conjugation) become F + because the F factor replicates during transfer. thus breaking the chromosome is transferred. an example of a class of genetic elements called episomes. like the chromosomes of specialized transducing phages. The F factor is thus. The integration of the F factor is believed to be mediated by short DNA sequences called IS elements. a site-specific recombination event. When an F+ donor cell conjugates with an F. A cell carrying a integrated F factor is called an Hfr. the F factor mediates the transfer of a chromosome of the Hfr cell to a recipient (F -) cell. Usually. The F factor can integrate into the host chromosome at any one of many sites by a mechanism that appears analogous to the integration of the chromosomke of a specialized transducing phage.BT0203 Genetics and Cytogenetics covalently inserted into the host chromosome like any other set of chromosomal genes. Department of Genetic Engineering . namely. mixing a population of F+ cells with a population of F . Thus. only a portion of the Hfr chromosome is transferred before the cells separate.

although different Hfr’s initiated transfer from different sites on the chromosome. A map distance of 1 minute corresponds to the length of the segment of the chromosome transferred in 1 minute during conjugation. with the circular strand being replicated in the donor cell and the displaced stand being replicated in the recipient cell as it is transferred. in Hfr by F – matings. Transfer is believed to be initiated by an endonucleolytic nick in one strand at a specific site (the “origin” of transfer) on the F factor. Chromosome transfer appears to proceed at a fairly constant rate. Thus. Rex Arunraj Assistant Prof. the recipient F – cell acquires a complete F factor (thus becoming an Hfr donor) only in those rare cases when an entire Hfr chromosome.coli chromosome. Thus. is transferred. the time interval between the transfer of any two markers (easily determined by interrupted mating experiments) is a good estimate of the physical distance separating the markers on the chromosome. The standard E. The transfer of a complete chromosome from an Hfr to an F – cell takes from 90 to 100 minutes. Department of Genetic Engineering . depending on the strain. as the standard unit for measuring linkage in E. as in F + by F – matings. Gene mapping by conjugation Subsequent studies with different Hfr strains revealed similar fixed transfer sequences. as in Hfr by F – matings.BT0203 Genetics and Cytogenetics The mechanism of transfer of DNA from a donor cell to a recipient cell during conjugation appears to be the same whether just the F factor is being transferred. The remaining part of the F factor. representing the time interval between the transfer of markers in interrupted mating experiments. coli linkage map is thus divided into minute intervals from 0 (arbitrarily set at the thrA gene) to 100 minutes on the basis of interrupted mating experiments. The 5’ end of the nicked strand is then transferred through the conjugation tube into the recipient cell. however. It has therefore proven convenient to use the minute. is the last segment of DNA to be transferred.cell prior to the sequential transfer of chromosomal genes. It is now clear that the F factor can integrate at many different sites in the circular E . with its integrated F factor. or the chromosome is being transferred. Because the origin of transfer is within the integrated F factor. coli. one portion of the F factor is transferred from an Hfr cell to an F . Transfer is believed to be coupled to rolling circle replication. and the site of integration determines the origin of transfer characteristic of a given Hfr.

for example. and gal+ will occur among thr+ leu+ str-r recombinants with percentage frequencies of 90. the lower its frequency among the recombinants. what will the frequencies of the donor (say azi-s. 80. Donor markers azi-s. T1-s. Department of Genetic Engineering . be identical to the plateau frequencies observed in the interrupted mating experiment. the Hfr H by F – cross. with the frequency of the marker decreasing as a function of its distance from the selected (thr+ and leu+) donor markers. Rex Arunraj Assistant Prof. Consider.lac+.BT0203 Genetics and Cytogenetics Linkage relationships can also be determined from uninterrupted mating experiments. T1-s. when a new mutation is identified. and 25. Thus. its approximate location is usually first determined by interrupted conjugation mapping. lac+. If thr+ leu+ str –r recombinants are selected and scored for the presence of other segregating markers by replicating. The farther a marker is from the selected donor marker (in the HfrH experiment. in fact. interrupted mating experiments are simpler and more direct. Although uninterrupted conjugation experiments of this type can be used to determine linkage relationships. The frequency will. and gal+ markers be among the recombinants? Will these donor markers all be present with the same frequency? The frequencies of these donor markers are observed to vary. 40. Its exact location is then usually determined by transduction mapping. The marker frequency gradient is caused by two major factors: (1) the approximately constant probability per unit time of spontaneous rupture of the conjugation tube and the chromosome and (2) the decreasing probability that any two donor markers will be incorporated into the recipient chromosome by a single pair of recombination events (incorporation of a donor fragment into a recipient chromosome always requires two recombination events) as the distance separating the two markers increases. respectively. thr+ and leu+). with the matings being allowed to proceed uninterrupted for 1-2 hours.