CAPE Biology Unit 2: Module 2

Gene technology

Terms used in gene technology:

• Gene technology – the use of DNA or genes to produce a useful product.

• Genetic engineering – the use of technology to change the genetic material of an organism. This may
involve the transfer of a gene from one species to another.
• Genome – the complete set of genetic material of a species.
• Recombinant DNA (rDNA) – DNA of an organism that has been changed or altered by genetic
engineering.
• Transgenic organism/Recombinant organism – an organism that has its DNA altered through the
insertion of new DNA from another species. These organisms are said to have recombinant DNA (rDNA) as
their nucleic acid.
• Genetically Modified Organism (GMO) – an organism that has its DNA changed or altered by gene
technology without insertion of new DNA from another species.

Role of restriction enzymes in bacterial cells

Restriction enzyme, also called restriction endonuclease, is an enzyme produced by bacteria that cuts DNA at
specific sites. In the bacterial cell, restriction enzymes cut foreign DNA, thus eliminating infecting organisms.
A bacterium uses a restriction enzyme to defend against viruses called bacteriophages. When a virus infects a
bacterium, it inserts its DNA or RNA into the bacterial cell so that new viruses can be replicated. However, the
bacterial restriction enzymes prevent viral replication by cutting the virus DNA or RNA into many pieces.
Different bacterial species make different restriction enzymes.

Diagram illustrating the role of restriction enzymes in a bacterium

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CAPE Biology Unit 2: Module 2
Gene technology

Role of restriction enzymes in genetic engineering

In genetic engineering, specific restriction enzymes are extracted from bacterial cells and used to cut plasmids
so that a desired gene (cDNA) can be inserted.

Restriction enzymes would usually make staggered cuts on the plasmid to form segments of DNA with sticky
ends. Sticky ends are the ends of a gene that can easily form hydrogen bonds with other DNA molecules. The
nucleotide sequences where restriction enzymes cut are called restriction sites.

a circular DNA that consists

of antibiotic resistant gene
found in bacteria.

Plasmid pBR322

Restriction enzymes making staggered cuts with sticky ends

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Gene technology

Steps in Recombinant DNA technology (Gene cloning)

Isolation of gene Insertion of gene into plasmid Insertion of plasmid into bacterium Synthesis

(i) Isolation of insulin gene (cDNA)

Reverse transcription DNA Polymerase

mRNA cDNA (SS) cDNA (DS)

Isolation of Human insulin gene

Evaluation:

Complete the table below:

Features Transcription Reverse transcription

Enzymes involved
TRANSCRIPTASE
Precursor molecule
DNA mRNA
Product
RNA single stranded cDNA

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CAPE Biology Unit 2: Module 2
Gene technology bacteria is quarentined
and the plasmid is
(ii) Insertion of gene into plasmid vector extracted. restriction
enzyme is used to cut
the restriction enzyme
after which ligase is used
to insert the insulin gene
in the plasmid.

cuts the plasmid

Insertion of gene into plasmid vector

(iii) Insertion of plasmid into bacteria

the plasmid vector is added to a

culture of bacteria and calcium ions
is added to increase the
permeability of cell walls so bacteria
would absob the plasmid and then
antibiotic is added to kill the bacteria
that hasnt absorbed the plasmid.

Insertion of plasmid into bacteria

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Gene technology

(iv) Cloning of bacteria

a fermenter is designed for the reproduction of bacteria and produce the insulin hormone
Cloning of bacteria

Evaluation: Genetic engineering

mesosome

cytoplasm
DNA

plasmid

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Gene technology

(a) Contrast the role of restriction enzymes in bacterial cells and genetic engineering.

In the bacterial cell, restriction enzymes cut foreign DNA, thus eliminating infecting organisms.
…………………………………………………………………………………………………………………
In genetic engineering, specific restriction enzymes are extracted from bacterial cells and used
…………………………………………………………………………………………………………………
to cut plasmids so that a desired gene (cDNA) can be inserted.
…………………………………………………………………………………………………………………

…………………………………………………………………………………………………………………

[2 marks]

(b) Explain the steps involved in the use of recombinant DNA technology for the production of human insulin
under the following headings:
(i) Gene isolation
(ii) Gene insertion into vector
in gene isolation the beta cells are extracted from the pancreas and mRNA is extracted from
…………………………………………………………………………………………………………………
those beta cells. reverse transcriptase is used to to convert the mRNA into single stranded cDNA
…………………………………………………………………………………………………………………
and the DNA polymerase is used to convert the single stranded cDNA into double stranded cDNA.
…………………………………………………………………………………………………………………

…………………………………………………………………………………………………………………
bacteria is quarentined the plasmid vector is added to a
…………………………………………………………………………………………………………………
and the plasmid is culture of bacteria and calcium ions
extracted. restriction is added to increase the
enzyme is used Type
to text
cut here
…………………………………………………………………………………………………………………
permeability of cell walls so bacteria
the restriction enzyme would absob the plasmid and then
after which ligase is used antibiotic is added to kill the bacteria
…………………………………………………………………………………………………………………
to insert the insulin gene that hasnt absorbed the plasmid.
in the plasmid.
…………………………………………………………………………………………………………………
a fermenter is designed for the reproduction of bacteria and produce the insulin hormone
…………………………………………………………………………………………………………………

[6 marks]

(c) How do plasmids act as a defense mechanism for bacteria?

PLASMID CONTAINS THE GENE WHICH CODES FOR THE DEVELOPMENT OF RESISTANT
…………………………………………………………………………………………………………………
AGAINST ANTIBIOTICS
…………………………………………………………………………………………………………………

[2 marks]

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CAPE Biology Unit 2: Module 2
Gene technology

Gene therapy

Gene therapy is the transfer of a gene into the cells of a patient in order to treat a disease. This is an example of
the genetic engineering (recombinant DNA technology).

Ex vivo gene therapy

In ex vivo gene therapy, cells are withdrawn from the patient and new DNA is inserted into the cells before the
cells are transferred back into the patient. This method is used to treat patients with SCID.

(i) Severe Combined Immunodeficiency Disease (SCID)

In the case of SCID, patients lack the enzyme Adenosine deaminase (ADA) that helps in the development of
cells of the immune system. This results in a weakened immune system.
• Cause – inheritance of a recessive gene that codes for the faulty version of the enzyme ADA.
• Effects
• Treatment: Ex vivo gene therapy in which stem cells from the bone marrow of an individual are
removed and the gene for the production of ADA is inserted into the stem cells by using retroviruses as
vectors.

Disadvantage: This treatment however has resulted in the development of leukemia in some individuals
because the retroviruses may insert the gene into an area in the DNA that affects cell division.

In vivo gene therapy

In in vivo gene therapy, vectors are inserted into a patient’s body to change the DNA of the cell. This method
was used to treat lung cancer and Cystic fibrosis.

(i) In vivo gene therapy for cystic fibrosis:

• Use of liposomes to insert the dominant genes for normal CFTR protein into the cells of the respiratory
tract.
Disadvantage: However, this was a short-lasting treatment since the few nasal cells that were
transformed had a short life span.
• Use of viruses to insert of dominant genes for normal CFTR protein into the cells of the respiratory tract.
Disadvantage: However, the individuals experienced unpleasant side effects due to infection by the
virus.

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Gene technology

Gene therapy

Types of gene therapy:

(i) Somatic gene therapy – the process of transferring a gene with the use of vectors into somatic cells. This
Only specific somatic cells of the body are targeted. SCID (stem cells) and cystic fibrosis (respiratory tract
cells) are treated by somatic gene therapy.

(ii) Germline gene therapy - the process of transferring a gene with the use of vectors into the embryo. Upon
cell division the gene would be transferred to all of the cells of the body of the organism, thus, germline
gene therapy targets all of the cells of the body.

Medical implications of genetic engineering:

Positive

• Large amounts of medical products can be made such as insulin, erythropoietin and tissue plasminogen
activator.
• Provides treatment options for disorders such as SCID, cystic fibrosis, cancer etc. through gene therapy

Negative

• Gene therapy treatments can lead to an allergic reaction or infection producing inflammation in the case
of cystic fibrosis.
• Only recessive genetic disorders are able to be treated by gene therapy
• Germline gene therapy is illegal in many countries. Genes may mutate resulting in the transfer of faulty
genes from one generation to the next.

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Gene technology

Agricultural implications of genetic engineering:

Positive

• Some crops such as maize and cotton have been modified to have the Bt toxin gene which enables them
to be more resistant to insect pests. Farmers growing Bt crops greatly reduce their cost for insecticides
and therefore lower the cost for crop production.
• Crops have been modified to be more resistant to herbicides and therefore this increases yield, reduce
cost of production and ultimately the cost of food
• Golden rice provides vitamin A to treat night blindness
• Improve shelf life

Negative

• Useful insects such as pollinators that feed on the pollen grains of the Bt crops may die.
• The pollen grains of GMOs can be transferred to closely related species and thus change their features.

Ethical and social implications of genetic engineering:

Some religions think that scientists are attempting to play as God. Society may reject the new product

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Gene technology

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Variation

Variation refers to the differences in individuals.

Genetic variation is variation due to the inheritance of genes.

Sources of genetic variation:

• Crossing over
• Random segregation
• Random fertilization
• Mutations

Environmental variation is variation due to environmental factors such as diet, sunlight, root space,
availability of nutrients etc. This type of variation is not inherited.

Mutations

Mutation is a random unpredictable change in the DNA. This may be caused by errors that can occur during
DNA replication, transcription or translation and may also be caused by radiation.

Types of mutations:

(i) Gene mutation - changes in the sequence of nucleotides in the DNA that makes up a gene.
(ii) Chromosome mutation – random change in the number or structure of chromosomes.

Types of gene mutations:

• Substitution mutation – one base is substituted for another. This often has no effect (silent mutation)
since the genetic code is degenerate meaning that each amino acid is coded by more than one triplet e.g.
GAA and GAG both code for the amino acid Leucine.

• Frame shift mutations (Point mutations) – occurs when a base pair is loss or added to DNA. This means
that the entire code would be read differently. Deletion is the removal of a base pair while Insertion is the
addition of a new pair of bases into DNA.

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CAPE Biology Unit 2: Module 2
Variation

Types of gene mutations:

Sickle cell anaemia – substitution mutation

Sickle cell anaemia is a disorder in which the red blood cells become sickle shape and thus carry less
haemoglobin.

• Development of sickle cell anaemia - substitution mutation in which the base Thymine in the amino acid
glutamine (CTC) is substituted by Adenine in valine (CAC) resulting in the production of abnormal Beta-
globins which causes the red blood cell to become sickled shape. The type of haemoglobin is called Hbs
haemoglobin. Offsprings that inherit this gene would also develop it.

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Variation

Sickle cell anaemia – substitution mutation

Development of sickle cell anaemia

• Signs/symptoms: tiredness, pain, fever, anaemia, kidney failure.

• Treatment/prevention: no cure, pain relievers, blood transfusion helps to increase the number of normal
red blood cells in the individual so as to relieve anaemia.

Inheritance of Sickle cell anaemia (co-dominance)

Genotype Phenotype

HbAHbA Normal haemoglobin

HbAHbS Sickle cell trait

HbSHbS Sickle cell anaemia

Table showing the inheritance of sickle cell anaemia

Types of Chromosome mutations

• Aneuploidy – chromosomal mutation in which one daughter cell gets both copies of a chromosome due to
non-disjunction. Non-disjunction is the failure of a pair of chromosomes to separate during anaphase.

Diagram showing non-disjunction during Meiosis

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Variation

Types of Chromosome mutations

• Aneuploidy - Down’s syndrome (trisomy 21)

Down’s syndrome is the inheritance of an extra copy of chromosome 21. Down’s syndrome is an example
of aneuploidy. The resulting zygote has three Chromosome 21. Children with Down’s syndrome have
slanted eyes, flattened nose, cheerful personality, heart defects, muscle weakness, mental retardation.

Diagram showing inheritance of Down’s syndrome

• Translocation – chromosomal mutation in which a section of one chromosome is broken off and joined to
the end of a different chromosome.

Translocation mutation (Reciprocal translocation - two way transfer)

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Variation

Types of Chromosome mutations

• Deletion – loss of a chromosome segment e.g. Cri-du-chat syndrome (cry of the cat) results due a loss of a
segment in chromosome 5. Infants develop intellectual disability, small head size, low birth weight etc.

• Duplication – addition of an identical chromosome segment.

• Inversion – a chromosome segment is flipped 1800.

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Variation

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Natural selection

Darwin’s Theory of Natural selection states that in the struggle for existence, organisms that are more highly
adapted to their environment would survive and reproduce.

Relationship between variation and natural selection

Variation allows some individuals to be more adapted to their environment so that they can survive and
reproduce e.g.

• Brown peppered moths (Biston betularia) have better camouflage in a brown woody forest than white
peppered moth. Therefore, due to variation in skin colour, the Brown peppered moths became more adapted
to their environment and survive and reproduced while the white peppered moths were easily killed by
predators.
• Antibiotic resistance – some bacteria have plasmids containing antibiotic resistant gene. Therefore, some
bacteria could survive and reproduce when exposed to antibiotics while others die.

Darwin’s Observations and Deductions

Observation 1: Organisms produce more offspring than are needed to replace the parents.

Observation 2: Natural populations tend to remain stable in size over long periods.

Deduction 1: There is competition for survival (a ‘struggle for existence’).

Observation 3: There is variation among the individuals of a given species.

Observation 4: Some of these variations are inherited.

Deduction 2 Individuals with characteristics that best adapt them to their environment are most likely to survive
and reproduce.

Deduction 3 If these characteristics can be inherited, then the offsprings will pass the characteristics on to their
offsprings

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Natural selection

Darwin’s conclusion

Darwin concluded that if this happened over a long period of time, then the characteristics of a species could
gradually change as better adapted individuals were more likely to survive and pass on their adaptations to their
offsprings. Gradually, the species would become better adapted to their environment.

Environmental pressures or selection pressures

Factors which affect a population are called environmental pressures or selection pressures which include:

• Disease
• Competition for food and shelter
• Predation
• Lack of sunlight, water
• Temperature changes

Organisms that have traits that allow them to overcome these environmental pressures are more highly adapted
to their environment while organisms that do not overcome these environmental pressures will die. Therefore,
certain traits are either favored and allow for survival and an increase in population (high natality rate) or
unfavoured and lead to death and a decrease in population (high mortality rate).

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Natural selection

Types of Natural selection

The distribution of traits across a population that is without environmental pressures/selection pressure would
have a normal bell curve of trait distribution. For example, if no selection pressure acted on plant height, then
most plants would be of an average height and fewer being extremely short or extremely tall.

Figure 1: The standard distribution of a trait across a population

However, when selection pressures act on a trait, this distribution can be altered resulting in stabilizing
selection, directional selection or disruptive selection.

Stabilizing selection

This is a type of natural selection in which selection pressures act against the two extremes of a trait within a
population. For example, if plant height (small, medium, large) was acted on by stabilizing selection, then the
selection pressures would act against small and large height while medium height would be favored.

Small Medium Large

Figure 2: The effect of stabilizing selection on trait distribution

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Natural selection

Directional selection

This is a type of natural selection in which one extreme of the trait distribution experiences selection pressures
against it. The result is that the population's trait distribution shifts towards the other extreme. For example:
giraffe necks. There was a selection pressure against short necks, since individuals with short necks could not
reach as many leaves on which to feed. As a result, the distribution of neck length shifted to favor individuals
with long necks. Another good example is with brown peppered moths (Biston betularia).

Short Medium Long

Disruptive Selection

In disruptive selection, selection pressures act against individuals in the middle of the trait distribution and
therefore individuals at the two extremes of the trait are favored. The result is a bimodal or two-peaked curve.
For example, in black bellied seed crackers (Pyrenestes ostrinus), selection pressure acted against medium sized
beaks since large beaks seed crackers were capable of cracking hard seeds while small beak seed crackers fed
on mostly soft seeds. There were very few medium sized beak seed crackers.

Small beaks Medium beaks Large beaks

The effect of disruptive selection on trait distribution
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CAPE Biology Unit 2: Module 2
Speciation

Speciation is the formation of a new species from an existing one.

Species is a group of organisms with similar morphology and physiology which can interbreed to produce
fertile offsprings.

To produce a new species from an existing one, the individuals must:

• become morphologically or physiologically different from the members of the original species
• no longer be able to breed with the members of the original species to produce fertile offsprings

Isolation

Isolation is the splitting apart of the new species group from the original group.

Types of isolation:

• Geographical isolation – occurs due to a physical barrier such as mountain, river etc.
• Reproductive isolation – occurs when the two groups are so different from each other that they can no
longer interbreed. This may be due to ecological, temporal or reproductive barriers.

Types of speciation:

1. Allopatric speciation – the formation of a new species from an existing population due to geographical
isolation e.g. tiger beetles that have been separated by a mountain range may become different species. - ----

Allopatric speciation in tiger beetles

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CAPE Biology Unit 2: Module 2
Speciation

Types of speciation

2. Sympatric speciation – the formation of a new species from an existing population living in the same
place. This may be due to changes in mating rituals, feeding habits, etc. that may cause a group to become
reproductively isolated due to ecological, temporal or reproductive barriers e.g.

(a) Ecological barriers – The apple flies laid their eggs on apples and hawthorns. As the eggs developed
into adults, they continued feeding on their designated food source but the two population of apple flies
did not interact so mating did not occur between flies of different food sources. This resulted in the
formation of apple flies and hawthorn flies.

Sympatric speciation by ecological barrier in apple and hawthorn flies

(b) Temporal barriers – species that breed during different times of day, different seasons, or different
times of the year would not be able to mix gametes and therefore become isolated e.g. eastern and
western spotted skunks. The Eastern spotted skunk mates in late winter while the
Western spotted skunk mates late summer.

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CAPE Biology Unit 2: Module 2
Speciation

Types of speciation:

2. Sympatric speciation

(c) Reproductive barriers:

Prezygotic reproductive barriers:
• Different courtship behaviour - A mallard drake will only mate with a female who displays
appropriate courtship behaviour.
• Mechanical problems with mating – one organism may be smaller than the other or have different
shapes or sizes of reproductive organs.
• Gamete incompatibility

Postzygotic reproductive barriers:

• Hybrid sterility – even if a zygote develops successfully the resulting hybrid cannot produce gametes
and so cannot reproduce e.g. mule.

(d) Polyploidy – a type of mutation in which a cell receives more than two sets of homologous
chromosomes. This occurs because of non-disjunction during mitosis. This occurs mostly in plants.

Normal mitosis: Polyploidy during mitosis

Diploid (2n) 8 chromosomes
Diploid (2n) 8 chromosomes

Non-disjunction

Diploid (2n) 8 chromosomes Tetraploid (4n) 16 chromosomes

The resulting tetraploid organism (4n) can only mate with other tetraploid organisms to produce fertile
offsprings and thus experience reproductive isolation.

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CAPE Biology Unit 2: Module 2
Speciation

Biological Species concept

The biological species concept states that a species is a group of organisms with similar morphology and
physiology which can interbreed to produce fertile offsprings.

Limitations of the biological species concept:

• Does not apply to organisms that reproduce asexually

• Cannot be used to classify extinct organisms that are only known through fossils, old bones or skin.

Natural selection and Speciation

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