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When the S. cerevisiae genome was sequenced, only about 40% of its predicted gen

ID: 66646 • Letter: W

Question

When the S. cerevisiae genome was sequenced, only about 40% of its predicted genes had been previously identified in forward genetic screens. This left about 60% of predicted genes with no known function, leading some to dub the genes fun (function unknown) genes.

1)

As an approach to understanding the function of a certain fun gene, you wish to create a loss-of-function allele. How will you accomplish this?

As an approach to understanding the function of a certain fun gene, you wish to create a loss-of-function allele. How will you accomplish this?

inversion and homologous recombination

2)

You wish to know under what conditions the gene expressed. How will you ascertain such information?

You wish to know under what conditions the gene expressed. How will you ascertain such information?

homologous recombination to create a knockout injection of dsRNA for RNA interference random transposable element insertion treat with a chemical mutagen

inversion and homologous recombination

2)

You wish to know under what conditions the gene expressed. How will you ascertain such information?

You wish to know under what conditions the gene expressed. How will you ascertain such information?

PCR and sequencing of the gnee RNAi knockdown transcriptional fusion with GFP gene homologous recombination to create a knockout

Explanation / Answer

Q1. Injection of dsRNA for RNA interference. RNA interference is a technique used for loss-of-function phenotype where the expression of protein is significantly reduced because of the destruction of mRNA or inability to translate it.

Q2. Transcriptional fusion with GFP gene. This GFP tag to the protein will allow the expression of this gene to be studied both microscopically and biochemically.

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