Whether the gene GAL4 or GAL80 genes can be considered as potential candidates for isolation of mutations that allow the constitutive expression of the target genes along with the characteristic of protein that disrupts the mutation. Introduction: Yeast genes have cis-acting elements upstream of their promoters and are similar to enhancers, which are called upstream activating sequences (UASs). UAS G regulates the several target genes involved in the galactose utilization.

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Gal4 is a transcription factor that activates transcription of galactose metabolism genes in yeast. These genes are ‘turned on’ when yeast cells need to metabolize galactose. To identify promoter sequences necessary for regulation of transcription of GAL1, reporter gene fusions were made and introduced into yeast cells. Deletions of GAL1 promoter were cloned upstream of LacZ gene. β-Galactosidase activity was measured in presence of galactose. Shown below is a representation of the results obtained. In the diagrams below (not to scale!): • Construct 1 contains ~ 130bp of the promoter, which is predicted to have all the predicted/putative proximal promoter elements (indicated by the solid boxes) needed to regulate transcription of GAL1.• The stippled box is the core promoter.• The arrow represents the transcriptional start site for the reporter gene Lac Z• Number of + signs represents level of transcription• Star represents a mutation in DNA sequence at that location (few nucleotides…

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Question

Chapter 17, Problem 8P

Summary Introduction

To determine:

Whether the gene GAL4 or GAL80 genes can be considered as potential candidates for isolation of mutations that allow the constitutive expression of the target genes along with the characteristic of protein that disrupts the mutation.

Introduction:

Yeast genes have cis-acting elements upstream of their promoters and are similar to enhancers, which are called upstream activating sequences (UASs). UAS_G regulates the several target genes involved in the galactose utilization.

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Gal4 is a transcription factor that activates transcription of galactose metabolism genes in yeast. These genes are ‘turned on’ when yeast cells need to metabolize galactose. To identify promoter sequences necessary for regulation of transcription of GAL1, reporter gene fusions were made and introduced into yeast cells. Deletions of GAL1 promoter were cloned upstream of LacZ gene. β-Galactosidase activity was measured in presence of galactose. Shown below is a representation of the results obtained. In the diagrams below (not to scale!): • Construct 1 contains ~ 130bp of the promoter, which is predicted to have all the predicted/putative proximal promoter elements (indicated by the solid boxes) needed to regulate transcription of GAL1.• The stippled box is the core promoter.• The arrow represents the transcriptional start site for the reporter gene Lac Z• Number of + signs represents level of transcription• Star represents a mutation in DNA sequence at that location (few nucleotides…

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