Lec 5 - Ch6.5

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1
How does Tetrad Analysis confirm recombination occuring at the four strand stage?
shows that in tetrads, two of the four chromatids contain a combination of genetic material from both parental strands, indicating a crossover event between homologous chromosomes
2
Explain how we know that DNA breaks and rejoins during recombination (3)
- Genetic material is exchanged between homologous chromosomes
- creates hybrid DNA strands
- Rejoining occurs at specific points during the process
3
What experiment shows DNA molecules break and rejoin during recombination
- Meselson and Weigle infected E coli with heavy and light isotopes of N
- Heavy is AB and light is ab
- Results show Ab with mostly heavy DNA which supports hypothesis
4
List out the steps of Recombination
1. Double strand break formation
2. Resection
3. Strand Invasion
4. Double Holliday Junction
5. Branch Migration
6. Holliday Junction Resolution
5
Step 1: Double Strand break formation
Spo11 protein breaks phosphodiester bonds of BOTH strands of ONE chromatid
6
Step 2: Resection
5′ ends of each broken strand are degraded by an EXONUCLEASE to create 3′ single-stranded tails
7
Step 3: Strand Invasion
- Dmc1 helps one single-strand tail invade a non-sister chromatid and forms stable heteroduplex
- Displacement loop (D-loop) from invaded chromatid is stabilized by single-strand binding protein
8
Step 4: Double Holliday Junction
- D-loop and other 3′ tail form a second heteroduplex
- DNA synthesis replaces the DNA degraded during formation of the 3′ tails
- DNA ligase reseals the DNA backbones
9
Step 5: Branch Migration
Heteroduplex region of both DNA molecules is lengthened
10
Step 6: Holliday junction Resolution
Holliday junctions are resolved by resolvase and ligase
11
why recombination events do not always result in crossing-over
- Invading strand is lengthened by DNA synthesis using blue strand as template
- Anticrossover helicase disentangles the invading strand from the nonsister chromatid preventing Holliday junction formation
12
What happens during mismatching in heteroduplexes? How is it resolved?
- Two strands of DNA don’t break and rejoin at the same location
- DNA repair enzymes eliminate mismatches and gene conversion occurs.
13
Gene conversion
the physical change of one allele in a heterozygote to the other
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