When the S.cerevisiae genome was sequenced and surveyed for possible genes, only about 40% of those genes had been previously identified in forward genetic screens.  This left about 60% of predcited genes with no known function, leading some to dub the genes fun (function unknown) genes.a)As an approach to understanding the function of a certain fun gene, you wish to create a loss of function allele.  How would you do this?b)You wish to know the physical location of the encoded protein product.  How would you obtain such information?

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Asked Nov 18, 2019
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When the S.cerevisiae genome was sequenced and surveyed for possible genes, only about 40% of those genes had been previously identified in forward genetic screens.  This left about 60% of predcited genes with no known function, leading some to dub the genes fun (function unknown) genes.

a)As an approach to understanding the function of a certain fun gene, you wish to create a loss of function allele.  How would you do this?

b)You wish to know the physical location of the encoded protein product.  How would you obtain such information?

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Expert Answer

Step 1
  • Loss of function allele can be created by transposon mediated insertional inactivation. The insertion of transposon into a gene can disrupt its reading frame. Such a disrupted gene expresses truncated polypeptides.
  • Two components are used for transposon mediated insertional inactivation: a plasmid carrying a reporter gene flanked by transposase recognition sequence and a DNA (deoxyribonucleic acid) segment containing the transposase gene. The separation of recognition sequence and transposase gene prevent the uncontrolled transposition of the reporter gene. The...

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