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Ch. 16 - Regulation of Gene Expression in Bacteria
Klug - Concepts of Genetics  12th Edition
Klug12th EditionConcepts of Genetics ISBN: 9780135564776Not the one you use?Change textbook
Chapter 16, Problem 23

A fellow student considers the issues in Problem 22 and argues that there is a more straightforward, nongenetic experiment that could differentiate between the two types of mutations. The experiment requires no fancy genetics and would allow you to easily assay the products of the other SOS genes. Propose such an experiment.

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Identify the two types of mutations being discussed in Problem 22. These are likely related to the SOS response, which is a bacterial DNA repair system activated under stress. Clarify whether the mutations affect the regulatory genes (e.g., lexA or recA) or the structural genes of the SOS system.
Understand the role of the SOS genes. The SOS response involves the activation of multiple genes, including those for DNA repair and error-prone polymerases. The products of these genes can be assayed to determine whether the SOS system is functioning.
Propose a nongenetic experiment to differentiate between the two types of mutations. For example, you could use a biochemical assay to measure the activity of proteins encoded by the SOS genes. This could involve testing for enzymatic activity (e.g., DNA polymerase activity) or detecting the presence of specific proteins using techniques like Western blotting.
Design the experiment to compare the activity of the SOS gene products in wild-type bacteria versus bacteria with the mutations. For instance, grow the bacteria under conditions that induce the SOS response (e.g., UV irradiation or DNA-damaging agents) and then measure the activity or presence of the SOS gene products.
Analyze the results to determine the type of mutation. If the SOS gene products are not produced or are nonfunctional, the mutation likely affects the structural genes. If the SOS gene products are produced but the response is not properly regulated, the mutation likely affects the regulatory genes (e.g., lexA or recA).

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Key Concepts

Here are the essential concepts you must grasp in order to answer the question correctly.

Types of Mutations

Understanding the different types of mutations, such as point mutations, insertions, deletions, and frameshift mutations, is crucial. Each type can affect gene function differently, leading to various phenotypic outcomes. Recognizing these distinctions helps in designing experiments to differentiate between mutations based on their effects on gene expression or protein function.
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Mutations and Phenotypes

SOS Response in Bacteria

The SOS response is a cellular mechanism in bacteria that is activated in response to DNA damage. It involves the expression of a set of genes that help repair DNA and can lead to increased mutation rates. Familiarity with the SOS genes and their products is essential for proposing experiments that assess the functionality of these genes under different conditions.
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Bacteria and Viral Chromosome Structure

Experimental Design in Genetics

Effective experimental design is fundamental in genetics to test hypotheses and differentiate between genetic and non-genetic factors. This includes selecting appropriate controls, determining the variables to measure, and choosing suitable assays. A well-structured experiment can provide clear insights into the mechanisms underlying mutations and their effects on cellular processes.
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Related Practice
Textbook Question

A marine bacterium is isolated and shown to contain an inducible operon whose genetic products metabolize oil when it is encountered in the environment. Investigation demonstrates that the operon is under positive control and that there is a reg gene whose product interacts with an operator region (o) to regulate the structural genes, designated sg. In an attempt to understand how the operon functions, a constitutive mutant strain and several partial diploid strains were isolated and tested with the results shown in the following table. Host Chromosome F' Factor Phenotype Wild type None Inducible Wild type reg gene from mutant strain Inducible Wild type Operon from mutant strain Constitutive Mutant strain reg gene from wild type Constitutive Draw all possible conclusions about the mutation as well as the nature of regulation of the operon. Is the constitutive mutation in the trans-acting reg element or in the cis-acting o operator element?

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Textbook Question

The SOS repair genes in E. coli are negatively regulated by the lexA gene product, called the LexA repressor. When a cell's DNA sustains extensive damage, the LexA repressor is inactivated by the recA gene product (RecA), and transcription of the SOS genes is increased dramatically. One of the SOS genes is the uvrA gene. You are a student studying the function of the UvrA gene product in DNA repair. You isolate a mutant strain that shows constitutive expression of the UvrA protein. Naming this mutant strain uvrAᶜ, you construct the diagram shown above in the right-hand column showing the lexA and uvrA operons:

Describe two different mutations that would result in a uvrA constitutive phenotype. Indicate the actual genotypes involved.

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Textbook Question

The SOS repair genes in E. coli are negatively regulated by the lexA gene product, called the LexA repressor. When a cell's DNA sustains extensive damage, the LexA repressor is inactivated by the recA gene product (RecA), and transcription of the SOS genes is increased dramatically. One of the SOS genes is the uvrA gene. You are a student studying the function of the uvrA gene product in DNA repair. You isolate a mutant strain that shows constitutive expression of the UvrA protein. Naming this mutant strain uvrAC, you construct the diagram shown above in the right-hand column showing the lexA and uvrA operons:

Outline a series of genetic experiments that would use partial diploid strains to determine which of the two possible mutations you have isolated.

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Textbook Question

The following figure depicts numerous critical regions of the leader sequence of mRNA that play important roles during the process of attenuation in the trp operon. A closer view of the leader sequence, which begins at about position 30 downstream from the 5' end, is shown below, running along both columns. Within this molecule are the sequences that cause the formation of the alternative hairpins. It also contains the successive triplets that encode tryptophan, where stalling during translation occurs.

Take a large piece of paper (such as manila wrapping paper) and, along with several other students from your genetics class, work through the base sequence to identify the trp codons and the parts of the molecule representing the base-pairing regions that form the terminator and antiterminator hairpins shown in the following figure.

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