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17 Cards in this Set
- Front
- Back
vulva
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the mating & egg-laying structure in hermaphrodites
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vulvaless (vul) and multivulval (muv)
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Two phenotypes associated with defects in vulval
development |
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vulvaless
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worms can’t lay eggs
so eggs hatch inside mom & eat their way out |
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multivulval
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multiple cells adopt
vulval cell fate |
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muv1(lf) --> vulval cell fate
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recessively
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muv1(gf --> vulval cell fate
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these mutants
act dominantly |
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how to perform an epistasis test?
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Using 2 mutants that give different phenotypes, generate double homozygous mutants and compare them to each single mutant
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What is a complementation test?
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Cross 2 homozygous mutants that give a similar phenotype and analyze the phenotype of the progeny
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Loss of function mutations = ________
Gain of function mutations = __________ vulvaless or multivulva |
Loss of function mutations = vulvaless
Gain of function mutations = multivulva |
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What would be a good general approach to identify genes that function downstream of let-23 in the VPCs?
Mutagenize let-23-/- and screen for Vul mutants Mutagenize let-23-/- and screen for Muv mutants Mutagenize let-23(gf)/+ and screen for Vul mutants Mutagenize let-23(gf)/+ and screen for Muv mutants |
Mutagenize let-23-/- and screen for Muv mutants
Mutagenize let-23(gf)/+ and screen for Vul mutants |
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A fifth gene in the pathway is identified, mek-2. Loss of function mutations
produce a vulvaless phenotype and gain of function mutations produce a multivulval phenotype. You think mek-2 encodes a protein that acts between LET-60 and MPK-1. Which double mutant combinations would allow you to determine that it acts downstream of LET-60 and upstream of MPK-1? mek-2(gf)/+; let-60(lf)/let-60(lf) and mpk-1(gf)/+; let-60(lf)/let-60(lf) mek-2(gf)/+; let-60(gf)/+ and mek-2(gf)/+; mpk-1(gf)/+ let-60(gf)/+; mek-2(lf)/mek-2(lf) and mek-2(gf)/+; mpk-1(lf)/mpk-1(lf) let-60(gf)/ let-60(gf); mpk-1(lf)/+ and let-60(gf)/ let-60(gf); mek-2(lf)/+ |
let-60(gf)/+; mek-2(lf)/mek-2(lf) and mek-2(gf)/+; mpk-1(lf)/mpk-1(lf)
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PCR
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use primers that will report on the presence of neo DNA
or on the absence of part of the wild-type gene |
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Southern
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use a probe that will report on the presence of neo DNA
or on the absence of part of the wild-type gene |
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Sequencing
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sequence the middle of the gene
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You generate a mouse knock-out construct so that a portion of the gene is
removed when it homologously recombines. You know that EcoRI sites flank the gene you knocked out. If you perform a Southern blot on EcoRI digested DNA from F3 mice, which probe will allow you to identify your homozygous mutant mice with out additional molecular analysis (i.e. PCR or sequencing)? a) probe against the portion of the wildtype gene that is removed b) probe against the neomycin resistance marker c) neither of these probes will avoid additional molecular analysis d) either of these probes will avoid additional molecular analysis |
probe against the portion of the wildtype gene that is removed
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maternal effect
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genotype of mom important
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zygotic genes
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genotype of embryo important
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