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Molecular Genetics LSM1102

S/N 1.

Lecture Topics (1) Reproduction and Chromosome Transmission (Cellular Division: Mitosis and Meiosis) (2) Gene Transfer/Mapping in Bacteria & Bacteriophages (3) Chromosome Organization and Structure (4) Recombination and Transposition Continual Assessment 1 (on Lectures 1-4) (10% of module scores)

Lecture hours 8 Wu Jinlu (Sem 1 & 2)

2.

(5) Molecular structure of DNA and RNA (6) DNA replication (7) Gene transcription and RNA processing (8) Translation of mRNA Continual Assessment 2 (on Lectures 5 8) (10% of module scores) 5-8)

8 He Yuehui (Sem 1) Liou Yih-Cherng (Sem 2)

3.

(9) Mendelian genetics + Sex Chromosomes and Sex Linkage (10) Modes of Inheritance and Pedigree Analysis (11) Variations to Mendelian Genetics, Penetrance, Expressivity Genetics Penetrance Expressivity, Pleotrophy and Linkage (12) Population Genetics Mutation and Selection, Fitness, Overdominance (13) Genetic Epidemiology Continual Assessment 3 (on Lecture 9 onwards) (10% of module scores)

10 Chew Fook Tim (Sem 1 & 2)

Laboratory sessions Practical 1


Extraction of genomic DNA from E. coli Basic bacterial genetic manipulation colony isolation and differential plating g p y p g

Practical 2
Agarose gel electrophoresis Extraction of plasmid DNA from E. coli Basic bacterial genetic manipulation grid plating

Practical 3
Bacterial Transformation Basic bacterial genetic manipulation replica plating

Dr. Wu Jinlu Dr. He Yuehui LS Lab 9 @MD4 Level 4

Practical P ti l 4
Observation of transformation and replica plating results Submission of laboratory reports (for Practicals 1-3) (up to 10% of module scores)

Practical 5
Observation of D Ob ti f Drosophila wild t hil ild types and mutants d t t Genetic Epidemiology collection of DNA sample, consent and ethics in research Genetic epidemiology collection of phenotypes by survey and objective testing Take Home Assignment - on Mendelian Genetics and Molecular Basis of Inheritance

Practical 6
Hypothesis testing in Mendelian genetics Take Home Assignment - on Variations to Mendelian Genetics

Practical (Tutorial) 7 + 8
Modes of Inheritance and Human Pedigree Analysis Hardy-Weinberg Equilibrium Population Genetics and Genetic Epidemiology (analysis) Take Home Assignment - on Population Genetics

Dr. Chew FT LS Lab 1&2@S1A Level 3

Text Book:
d d Genetics - Analysis and Principles, 2nd or 3rd Edition

Robert J. Brooker

Reference
Concepts of Genetics: International Edition, 9/E, ISBN-10: 0321540980 Principles of Genetics, 4th Edition ISBN: 978-0471-69939-2

Lecturers: L t
Coordinator (for both Semesters I and II) Dr Chew Fook Tim (Department of Biological Sciences) Email: dbscft@nus.edu.sg Tel: 6516 1685 Block S3, Level 2, Functional Genomics Laboratories, Room 5 Dr Wu Jinlu (Department of Biological Sciences) (Sem I and II) Email: dbswjl@nus.edu.sg Tel: 6516 8476; S1A-2-27 Dr He Yuehui (Department of Biological Sciences) (Sem I) Email: dbshy@nus.edu.sg Tel: 6516 2716 Dr Liou Yuh-Cherng (Department of Biological Sciences) (Sem II) Email: bchlyc@nus.edu.sg Tel: 6516 7711
Module

Wu Jinlu dbswjl@nus.edu.sg j@ g Tel: 65168476; S1A-L2-27

Keep yourself informed _ http://ivle.nus.edu.sg/lms/

IDEA?

Chromosome
1.Separation 1 Separation and Transmission (eukaryotes) 2. Separation and Transmission (Prokaryotes) (P k t ) 3.Organization and Structure 3O i ti d St t 4.Recombination 4 Recombination and Transposition

CHAPTER 3
REPRODUCTION AND CHROMOSOME TRANSMISSION
Genetics: analysis & principles Robert J. Brooker

3.1 GENERAL FEATURES OF Chromosomes CHROMOSOMES


y, Biochemically, chromosomes are composed of p DNA, which is the genetic material Proteins which Proteins, provide an organized structure
Greek khrma colour + sma body

Eukaryotic Chromosomes Are Inherited in Sets


Most eukaryotic species are diploid Have two sets of chromosomes in somatic cells Members of a pair of chromosomes are called homologues The two homologues form a homologous pair The two chromosomes in a homologous pair Are nearly identical in size y Have the same banding pattern and centromere location Have the same genes But not necessarily the same alleles

Genes, Alleles and Chromosome

Dominant Recessive
The physical location of a gene on a chromosome is called its locus.

3.2 CELLULAR DIVISION-MITOSIS


G0 phase

Interphase (G1, S, G2)


Chromosomes are decondensed B th end of this By the d f thi phase, the chromosomes have already replicated The centrosome divides

http://fig.cox.miami.edu/~cmallery/150/cells/c7x22centrosome.jpg

Mitosis is subdivided into five phases (PPMAT) Prophase Prometaphase Metaphase M t h Anaphase Anaphase Telophase

Prophase
Nuclear envelope dissociates into smaller vesicles Centrosomes separate to opposite poles The mitotic spindle apparatus is formed

Prometaphase
Kinetochore microtubules grow from the two poles Spindle fibers interact with the sister chromatids h tid The two kinetochores on a pair of sister chromatids are attached to kinetochore MTs on opposite poles

Metaphase
Pairs of sister g chromatids align themselves along a p plane called the metaphase plate Each pair of chromatids is attached to both poles by kinetochore microtubules

Anaphase
The connection holding the sister chromatids together is broken Each chromatid, now an individual chromosome, is linked to only one pole

pull push h

Bold arrows show the force direction

Telophase and Cytokinesis


Chromosomes reach their respective poles and decondense Nuclear membrane reforms to form two separate nuclei I most cases, In t mitosis is quickly followed by f ll db cytokinesis

Animation
mitosis
http://www.youtube.com/watch?v VlN7K1 9QB0&feature related http://www.youtube.com/watch?v=VlN7K1-9QB0&feature=related http://highered.mcgraw-hill.com/sites/0072835125/student_view0/index.html

3.3 MEIOSIS
Meiosis I and II Each of these is subdivided into Prophase Prometaphase p Metaphase A Anaphase h p Telophase

Prophase I
Prophase I is further subdivided into periods known as (LZPDD)

Leptotena (thin and delicate) Leptotena Zygotena (conjugation) Pachytena (thick or fat) Diplotena (two fold or double) Diplotena Diakinesis (thin and delicate)

A tetrad

A physical exchange of chromosome pieces

Spindle apparatus complete Chromatids attached via kinetochore microtubules ki t h i t b l

Metaphase I
Bivalents are organized along the metaphase plate P i of sister chromatids Pairs f i t h tid are aligned in a double row, row rather than a single row (as in mitosis) Furthermore A pair of sister chromatids is linked to one of the poles And the homologous p g pair is linked to the opposite pole

Figure 3.13

Figure 3.17

During metaphase I, the bivalents can align themeselves in two different ways

Independent assortment of the R/r and Y/y alleles

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Independent assortment refers to the random


arrangement of p g pairs of chromosomes. The diagram below g shows four possible arrangements of chromosomes during metaphase 1 from an individual that has 6 total chromosomes. chromosomes Suppose that the pink chromosomes are those that the individual inherited from its mother and the blue colored ones were inherited from its father. For each chromosome pair, the chromosome that is on the left (maternal or paternal) is determined randomly. As can be , g p seen, there are several alignment possibilities.

Down syndrome

Trisomy

The two pairs of sister chromatids separate from each other However, th connection that H the ti th t holds sister chromatids together does not break

Sister chromatids reach their respective poles and decondense Nuclear envelope reforms to produce N l l f t d two separate nuclei

Meiosis II

Mitosis vs Meiosis_Summary
Mitosis produces two diploid daughter cells Meiosis produce four haploid daughter cells Mitosis produces daughter cells that are genetically identical Meiosis produces daughter cells that are not genetically identical The daughter cells contain only one homologous chromosome from each pair

Animation
Meiosis M i i I & II
http://www.youtube.com/watch?v=D1_-mQS_FZ0&feature=related http://highered.mcgraw-hill.com/sites/0072835125/student_view0/index.html

http://www.youtube.com/watch?v=R_LUJSqeSrI

Can you tell the stage of cells Can o (image in the right hand) What do the different colors represent for?

Q: A diploid cell has eight


chromosomes, four per set. In the h f t I th following diagram, what phase of mitosis, meiosis I or meiosis II, is this cell in?
The cell is in metaphase of meiosis II. You can tell because the chromosomes are lined up along the metaphase plate, and it plate has only four pairs of sister chromatids. If it were mitosis, the cell would have eight pairs of ll ld h i ht i f sister chromatids.

A1:

Q2:

A cell is diploid and contains three chromosomes per set. Draw the arrangement g of chromosomes during metaphase of mitosis, and metaphase I and II of meiosis.

A2:

Critical thinking g /application Infertility of mules


Mules are the result of a c oss between cross bet ee a horse ( o se (2n = 64) mother and donkey (2n = 62) father

http://www.horsegroomingsupplies.com/horseforums/attachments/horse-chat/45122-mulesdressagemule.jpg

What we have not touched/known


1. Division of intracellular organelles 2. Cell cycle control 3. Mechanics of cell division 4..

Ordinary bull or incredible hulk _Muscles, Mutations, and Myostatin

3.4 Sex Chromosome/Determination


Humans have 46 chromosomes

44 autosomes 2 sex chromosomes h

Males contain one X and one Y chromosome


(heterogametic)

Females have two X chromosomes


(homogametic)

The Y chromosome determines maleness

Figure 3.20

Contains many X-linked genes

Involved in o ed antibody production

Y-linked gene Follows a pseudoautosomal pattern of inheritance Necessary for proper male development l d l t

In some insects,
Males are XO and females are XX

In other insects (fruit fly, for example)


Males are XY and females are XX

The Y chromosome does not determines maleness Rather, it is the ratio between the X chromosomes and the number of sets of autosomes (X/A)
If X/A = 0.5, the fly becomes a male If X/A = 1.0, the fly becomes a female If 0.5 <X/A <1.0, it develops as an intersex X/A 1.0,

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X/A ratio and sex determination


Drosophila M M.

Q: Assuming that such a fly would be viable, what would


be th b the sex of a fruit fly with the following chromosomal f f it fl ith th f ll i h l composition A. A One X., and two sets of autosomes X B. Two X., one Y., and two sets of A. C. Two X, two Y, and four sets of A, D. Four X, two Y, and four sets of A,

The sex chromosomes are designated Z and W to distinguish them from the X and Y chromosomes of mammals Males contain two Z chromosomes
Hence, they are homogametic

Females have one Z and one W chromosome


Hence, they are heterogametic

and some fish

3-80

Males are known as the drones


They are haploid Produced from unfertilized eggs

Females include the worker bees and queen bees


They are diploid Produced from fertilized eggs

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Why does sex exist?

http://sciencenow.sciencemag.org/cgi/content/full/2000/1017/3

Inheritance and biogenesis of organelles in the secretory pathway Martin Lowe and Francis A Barr A. The inheritance of organelle genes and Th i h it f ll d genomes: patterns and mechanisms Jianping X Ji i Xu