Prescott's Microbiology
Prescott's Microbiology
10th Edition
ISBN: 9781259281594
Author: Joanne Willey, Linda Sherwood Adjunt Professor Lecturer, Christopher J. Woolverton Professor
Publisher: McGraw-Hill Education
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Chapter 15, Problem 5CHI
Summary Introduction

The gene expression in yeast offers many advantages in producing proteins as compared to other eukaryotic organisms. There are many expression systems in the severalgenera ofyeasts. For example, Saccharomyces, Pichia and Yarrowia. Expression vector is used in this process. In most of the cases, the vectors that integrate with the host chromosome is widely used.

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Searching the yeast Saccharomyces cerevisiae genome, researchers found approximately 4,000 DNA sites with a sequence which could potentially bind the yeast transcription factor GAL4.  GAL4 activates the transcription of galactose genes.  Yet there are only 10 GAL4-binding sites which control the genes necessary for galactose metabolism.  The GAL4 binding sequence is CGGAT#AGAAGC*GCCG, where # is T, C or G, and * is C or T. In one chromatin immunoprecipitation experiment (ChIP), yeast growing on galactose were lysed, and subjected to cross-linking reagents which cross-linked transcription factors and activators to DNA. Next the DNA was sheared into small fragments, and antibodies to GAL4 were added.  These antibodies coprecipitated the GAL4 and the DNA it was cross-linked to.  The cross-linking was then chemically reversed, and the DNA was isolated, cloned into a library of plasmids and sequenced.  Results showed that only 10 different DNA sequences had GAL4 bound.  Since the…
Explain how the following mutations would affect transcription of the yeast GAL1 gene in the presence of galactose. (a) A deletion within the GAL4 gene that removes the region encoding amino acids 1 to 100. (b) A deletion of the entire GAL3 gene. (c) A mutation within the GAL80 gene that blocks the ability of Gal80 protein to interact with Gal3p. (d) A deletion of one of the four UASG elements upstream from the GAL1 gene. (e) A point mutation in the GAL1 core promoter that alters the sequence of the TATA box.
Debes et al recently described how aging in humans, mice, nematodes, and other eukaryotes is associated with an increase in the average speed of transcriptional elongation by RNA polymerase II. Overexpression of some proteins that decreased PolII elongation speed extended lifespan in the fly Drosophila. For each of the following proteins, predict whether overexpression of that protein (assuming all other cellular components are normal) would likely reduce transcriptional speed, and briefly justify your answer.  a)    Mediator proteinsb)    Histone proteinsc)    Insulator binding proteins
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Mitochondrial mutations; Author: Useful Genetics;https://www.youtube.com/watch?v=GvgXe-3RJeU;License: CC-BY