PRESCOTT'S MICROBIOLOGY
PRESCOTT'S MICROBIOLOGY
11th Edition
ISBN: 2818440045677
Author: WILLEY
Publisher: MCG
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Chapter 18, Problem 2AL
Summary Introduction

EMSA stands for electrophoretic mobility shift assay and it is the most common method used to study DNA-protein interactions. This method is also termed as a gel shift assay. This is because the DNA binding protein is added to the purified target DNA mixture and its mobility is slowed down using an agarose gel. The DNA-protein complex is found to be shifted to a higher molecular weight when compared to the DNA without the protein. Usually, the magnitude of the shift is related to the DNA-protein ratio. Thus, the largest shift would be seen when sufficient protein is added.

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The amino acid asparagine is synthesized from aspartic acid by the enzyme asparagine synthetase (AS). In the previous problem you proposed a model for how this gene could be regulated. Suppose that you carry out an experiment to test your model. To do this you cut out the regulatory sequences upstream of the gene and fuse it to a gene for green fluorescent protein (GFP). Now you can visually observe when the gene is activated. You insert this engineered gene into a host cell and look for GFP expression. You discover some mutants that have different expression levels of GFP and call them GFP1- and GFP2-. The expression levels of GFP are given below. Cell GFP expression Wild type 100 GFP1- 50 GFP2- 0 Propose an explanation for these results based on your model. In other words, what was mutated and how? This answer should include whether the mutation is (view links for more information): dominant or recessive https://www.ncbi.nlm.nih.gov/books/NBK21578/#A1877 in a cis…
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