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Genetic Analysis I > 6.5 and 6.6 > Flashcards

6.5 and 6.6 Flashcards

1
Q

Recombination prefers homologous because

A

no loss of nucleotides at repair site

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2
Q

Homologous Recombination used in

A

Meiosis and DNA Damage Response

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3
Q

Homologous Recombination specific for

A

Double Stranded Breaks (DSB).

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4
Q

Homologous Recombination exchanges

A

similar/identical genetic information across two separate strands of DNA

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5
Q

Meselson-Weigle Experiment

A

Bacteria (Lambda) was grown/maintained in
“Light” medium

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6
Q

Lambda was chosen bc

A

Half DNA by weight so density of the whole virus reflects the density of its DNA

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7
Q

Meselson-Weigle proved

A

crossing over also involves breakage and reunion of DNA molecule

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8
Q

Yeast as a Model Organism

A

*Cheap
*Abundant
*Fast to grow
*Eukaryotic
*Easy to Genetically Manipulate
*Can be studied as Haploid or Diploid
–Yeast can undergo Meiosis

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9
Q

Yeast

A

has help in the understanding of recombination

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10
Q

Spo11

A

protein that makes a double strand break on on of the chromatids by cleaving the phosphodiester bond

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11
Q

Exonuclease

A

enzyme that removes nucleotides from an end of a DNA molecule

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12
Q

Dmc1

A

takes 3’ tail and promotes invasion into homologous chromatid – forms heteroduplex

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13
Q

Heteroduplex

A

region of double stranded DNA in which two strands have nonidentical sequences
Formed during crossing-over
Held by hydrogen bonds

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14
Q

D-loop

A

Structure where the two strands of a double stranded DNA molecule are separated for a stretch and held apart by a third strand of DNA

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15
Q

D-loop stabilized by

A

binding of replication protein A (RPA)

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16
Q

Spo11 is highly

A

conserved

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17
Q

D-Loop connects with other 3’ tail to form

A

2nd heteroduplex

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18
Q

Holliday junctions

A

interlocked regions of two sister chromatids in recombination intermediates

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19
Q

Heteroduplex Regions

A

Expand outwards on both chromatids

20
Q

Resolvase

A

enzyme that breaks and joins DNA strands at Holliday junctions to separate nonsister chromatids during crossing-over

21
Q

Anticrossover helicase

A

helps disentangle the invading strand thus interrupting Holliday junction formation and preventing crossing-over

22
Q

If a cell were to respond to a DNA damage
event during homologous recombination,
where would there most likelybe an error?

A

Heteroduplex

23
Q

Branch migration

A

Holliday juncations move away from each other and enlarge the heteroduplx region between them

24
Q

exonuclease produces

A

two 3’ single-stranded tails

25
Q

Homologous Recombination steps

A

*Step 1: Spo11 causes DSB
*Step 2: Exonuclease degrades 5’end of breaks.
*Step 3: Strand invasion – 1st heteroduplex
*Step 4: Holliday Junctions – 2nd heteroduplex
*Step 5: Expansion of heteroduplex regions
*Step 6: Resolving Holliday Junctions

26
Q

Spo11 prefers to cleave

A

particular genomic sites, resulting in hot spots for cross-over

27
Q

Gene Conversion

A

Where one allele is changed to another allele during recombination as a result of heteroduplex formation and mismatch repair during recombination

28
Q

Mitotic cells do not make

A

Spo11 (no homologous recombination

29
Q

Mitotic recombinantion occurs bc

A

damage by environment agents that result in double strand breaks or single strand nicks in DNA

30
Q

Tetrad analysis

A

alleles segregate equally into gametes

31
Q

Gene conversion produces

A

tetrads will not segregate equally (breaks Mendel’s first low)

32
Q

Some organisms find it useful to have

A

site-specific recombination

33
Q

site-specific recombination is due to

A

recombinases

34
Q

site-specific recombination occurs only DNA target sites that are

A

less 200 base pairs long

35
Q

site-specific recombination is much

A

similar that homologous recombination

36
Q

Integration

A

insertion of one DNA molecule into another

37
Q

Cre-Recombinase

A

Recognizes LoxP sequences to perform recombination

38
Q

system becoming more frequently used in mouse-based research

A

FLP-FRT

39
Q

FLP-FRT system

A

using flippase (FLP) recombinase, derived from the yeast Saccharomyces cerevisiae
FLP recognizes a pair of FLP recombinase target (FRT) sequences that flank a genomic region of interest

40
Q

Two types of site specific recombination

A

FLP-FRT

41
Q

Uses of site specific recombo

A

turn on or off expression of a specific gene

42
Q

Clustered Regularly
Interspaced Short
Palindromic Repeats

A

CRISPR

43
Q

CRISPR

A

Prokaryotic Anti-viral
defense mechanism

44
Q

CRISPR/Cas9

A

induce double strand breaks at almost any position in the genome

45
Q

CRISPR/Cas9 allows for

A

edit genomes in vicinity of the breakage

Genetic Analysis I (9 decks)

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