Chapter 6
description
Transcript of Chapter 6
![Page 1: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/1.jpg)
31 January, 2 February, 2005
Chapter 6
Genetic Recombination in Eukaryotes
Linkage and genetic diversity
![Page 2: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/2.jpg)
Overview• In meiosis, recombinant products with new combinations of
parental alleles are generated by:– independent assortment (segregation) of alleles on nonhomologous
chromosomes.– crossing-over in premeiotic S between nonsister homologs.
• In dihybrid meiosis, 50% recombinants indicates either that genes are on different chromosomes or that they are far apart on the same chromosome.
• Recombination frequencies can be used to map gene loci to relative positions; such maps are linear.
• Crossing-over involves formation of DNA heteroduplex.
![Page 3: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/3.jpg)
Overview
![Page 4: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/4.jpg)
Recombination Detailed
![Page 5: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/5.jpg)
Independent Assortment
![Page 6: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/6.jpg)
Independent assortment (2)• For genes on different (nonhomologous) pairs of chromosomes,
recombinant frequency is always 50%
A/A ; B/B a/a ; b/b
A/a ; B/b
¼ A ; B P
¼ A ; b R
¼ a ; B R
¼ a ; b P
A/A ; b/b a/a ; B/B
A/a ; B/b
¼ A ; B R
¼ A ; b P
¼ a ; B P
¼ a ; b R
50%
recombinants
![Page 7: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/7.jpg)
Testcross of a Dihybrid
![Page 8: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/8.jpg)
Self Cross of a Dihybrid
![Page 9: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/9.jpg)
![Page 10: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/10.jpg)
![Page 11: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/11.jpg)
Independent assortment: multiple loci
• Calculations can be made for any gene combination using predicted outcomes at single loci and the product rule
P1 A/a ; B/b ; C/c ; D/d P2 a/a ; B/b ; C/c ; D/D
# gametes P1 2 x 2 x 2 x 2 = 16
# gametes P2 1 x 2 x 2 x 1 = 4
# genotypes in F1 2 x 3 x 3 x 2 = 36
# phenotypes in F1 2 x 2 x 2 x 1 = 8
Frequency of
A/– ; B/– ; C/– ; D/–
½ x ¾ x ¾ x 1 = 9/32
![Page 12: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/12.jpg)
Crossing-over • Breakage and rejoining of homologous
DNA double helices• Occurs only between nonsister chromatids
at the same precise place• Visible in diplotene as chiasmata• Occurs between linked loci on same
chromosome– cis: recessive alleles on same homolog (AB/ab)– trans: recessive alleles on different homologs
(Ab/aB)
![Page 13: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/13.jpg)
![Page 14: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/14.jpg)
![Page 15: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/15.jpg)
![Page 16: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/16.jpg)
![Page 17: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/17.jpg)
Linkage maps
• RF is (60+50)/400=27.5%, clearly less than 50%
• Map is given by:
# observed
140
50
60
150
A B
27.5 m.u.
![Page 18: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/18.jpg)
![Page 19: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/19.jpg)
![Page 20: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/20.jpg)
Trihybrid testcross
• Sometimes called three-point testcross
• Determines gene order as well as relative gene distances
• 8 categories of offspring– for linked genes, significant departure from
1:1:1:1:1:1:1:1
• Works best with large numbers of offspring, as in fungi, Drosophila
![Page 21: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/21.jpg)
Trihybrid testcross example
v (vestigial) v+ (long) wings
b (black) b+ (gray) body
p (purple) p+ (red) eyes
v+ v+ . b b . pp x vv. b+ b+ . p+ p+
v/ v+. b/ b+. p/ p+
![Page 22: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/22.jpg)
v/ v+. b/ b+. p/ p+ x v/v.b/b.p/p
Progeny:
v b+ p+ 580 Parental
v+ b p 592 Parental
v b p+ 45
v+ b+ p 40
v b p 89
v+ b+ p+ 94
v b+ p 3
v+ b p+ 5
![Page 23: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/23.jpg)
v.b and v+.b+ 45 + 40 + 89 + 94 = 268 / 1448 = .185
v.p and v+.p+ 89 + 94 + 3 + 5 = 191 / 1448 = .132
b.p+ and b+.p 45 + 40 + 3 + 5 = 93 / 1448 = .064
v 13.2 p 6.4 b
Why do 13.2 + 6.4 = 19.6 instead of 18.5?
![Page 24: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/24.jpg)
The failure to count double crossover gametes as v.b recombinants results in an underestimation of the v.b distance. A better v.b number would be 268 + 3 + 3 + 5 + 5 = 284 / 1448 = 19.6. Aaah!
In general, to minimize the effect of double crossovers, it is necessary to measure a number of small RF distances and sum to larger distances.
![Page 25: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/25.jpg)
Interference
• Crossing-over in one region of chromosome sometimes influences crossing-over in an adjacent region
• Interference = 1 – (coefficient of coincidence)
• Usually, I varies from 0 to 1, but sometimes it is negative, meaning double crossing-over is enhanced
tsrecombinan double expected #
tsrecombinan double observed # c.o.c. =
![Page 26: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/26.jpg)
In the previous example, expect (.132 x .064 = .0084)(1448) =
12 double crossovers, but saw 8, so I = 1-(8/12) = 4/12 = 1/3.
In regions where double crossovers are forbidden, observed = 0, so I = 1.
What do you think negative interference means?
![Page 27: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/27.jpg)
Genetic maps
•Useful in understanding and experimenting with the genome of organisms
•Available for many organisms in the literature and at Web sites
•Maps based on RF are supplemented with maps based on molecular markers, segments of chromosomes with different nucleotide sequences
![Page 28: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/28.jpg)
![Page 29: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/29.jpg)
Chi-square test • Statistical analysis of goodness of fit
between observed data and expected outcome (null hypothesis)
• Calculates the probability of chance deviations from expectation if hypothesis is true
• 5% cutoff for rejecting hypothesis– may therefore reject true hypothesis– statistical tests never provide certainty, merely
probability
![Page 30: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/30.jpg)
Chi-square application to linkage• Null hypothesis for linkage analysis
– based on independent assortment, i.e., no linkage
– no precise prediction for linked genes in absence of map
for all classes
• Calculated from actual observed (O) and expected (E) numbers, not percentages
∑ −=
E
EO 22 )(χ
![Page 31: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/31.jpg)
What is E?
For the AbBb testcross example from the text, E is not just 125 for each of the genotypes, as there may be allele effects on viability. Instead, get expected values from the data:
A aB 142 112 254
b 113 133 246
255 245
So, E (AB) would be (255/500) x (254/500) x 500 = 129.54
Other E are found similarly, and Chi-square is 4.97
![Page 32: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/32.jpg)
![Page 33: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/33.jpg)
Mechanism of meiotic crossing-over
•Exact mechanism with no gain or loss of genetic material
•Current model: heteroduplex DNA–hybrid DNA molecule of single strand from each of two nonsister chromatids–heteroduplex resolved by DNA repair mechanisms
•May result in aberrant ratios in systems that allow their detection
![Page 34: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/34.jpg)
![Page 35: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/35.jpg)
![Page 36: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/36.jpg)
Recombination within a gene• Recombination between alleles at a single
locus• In diploid heterozygous for mutant alleles of
the same gene, recombination can generate wild-type and double mutant alleles
• Rare event, 10-3 to 10-6, but in systems with large number of offspring, recombination can be used to map mutations within a gene
a1/a2 a+ and a1,2
![Page 37: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/37.jpg)
Assignment: Concept map, solved problems 1 and 2, all other basic and challenging problems
Continue with PubMed section of the Web tutorial.
![Page 38: Chapter 6](https://reader033.fdocuments.net/reader033/viewer/2022051620/5681399d550346895da13883/html5/thumbnails/38.jpg)