Microbiology: An Evolving Science (Fourth Edition)
Microbiology: An Evolving Science (Fourth Edition)
4th Edition
ISBN: 9780393615098
Author: John W. Foster, Joan L. Slonczewski
Publisher: W. W. Norton & Company
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Chapter 10.5, Problem 1TQ
Summary Introduction

To review:

The results of fusion of a promoter for an SOS response gene with the luciferase open reading frame.

Introduction:

The class of oxidative enzymes that produces bioluminescence is defined as luciferase. They have wide applications in the field of biotechnology. They are used as reporter genes isolated from the firefly Photinuspyralis. When luciferase acts on the appropriate luciferin substrate, light is emitted.

 

The reporter genes are nucleic acid sequences that encode products that can be detected in the transformed tissues. They are often used as indicators of whether a specific gene has been expressed in the organism or not. The genes that code for luciferase can serve as reporter genes as their expression is easy to monitor. Luciferase possesses some features that make them useful particularly as a reporter molecule for biosensing.

It can be used as a real-time biosensor for environmental stress response. RecA (recombination A) protein that is stimulated by the single-stranded DNA (deoxyribonucleic acid) is involved in the inactivation of the repressor (LexA) of SOS response genes, which induce the response. The fusion of the luciferase gene with a recA promoter will show an increase in the fluorescence after the irradiation by ultraviolet light by putting the culture into a spectrophotometer.

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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? Your answer should include whether the mutation is (see links for more information): dominant or recessive https://www.ncbi.nlm.nih.gov/books/NBK21578/#A1877…
Which of the following statements is/are TRUE for the promoters? The Pribnow box and TATA box are found in the prokaryotes and eukaryotes, respectively. For both prokaryotes and eukaryotes, the first ribonucleotide attaches to the TSS region. The -35 element in prokaryotes is called the Up element in eukaryotes. In prokaryotes, the Pribnow box is located 25 bases before TSS.
How does tryptophan regulates its own production? a. When tryptophan levels are high, it binds to the trp repressor, which changes its shape and binds to the operator sequence in the trp operon. b. When tryptophan levels are high, it binds to the trp repressor, which changes its shape and binds to the promoter sequence in the trp operon. c. When tryptophan levels are high, it binds to the trp repressor, which changes its shape and falls off from the operator sequence in the trp operon. d. When tryptophan levels are high, it binds to the trp repressor, which changes its shape and falls off from the promoter sequence in the trp operon.
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Mitochondrial mutations; Author: Useful Genetics;https://www.youtube.com/watch?v=GvgXe-3RJeU;License: CC-BY