Genetics: Analysis and Principles
6th Edition
ISBN: 9781259616020
Author: Robert J. Brooker Professor Dr.
Publisher: McGraw-Hill Education
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Chapter 15, Problem 6EQ
As described in Chapter 21, an electrophoretic mobility shift assay (EMSA) can be used to determine if a protein binds to a segment of DNA. When a segment of DNA is bound by a protein, its mobility will be retarded, and the DNA band will appear higher in the gel. In an EMSA whose results are shown below, a cloned gene fragment that is
in length contains a regulatory element that is recognized by a transcription factor called protein X. Previous experiments have shown that the presence of hormone X results in transcriptional activation by protein X.
Explain the action of hormone X.
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GR and PPAR are transcription factors that bind to GRE and PPARE sequences respectively and activate transcription of genes. A reporter cell line is created in which the the green fluoresecent protein (GFP) is controlled by a GRE sequence and the pink fluorescent protein mCherry is under control of a PPARE sequence. If the gene for GR is introduced into the reporter cell line, the cells produce a green color. Chimeric proteins are created in which the DNA Binding Domains (DBD) and Activation Domains (AD) of the transcription factors are introduced into various cell lines. Match the following cell-types with the fluorescent color(s) you would expect the cells to produce.
In cells, certain hormones, such as epinephrine, have the ability to raise the concentration of cAMP. The CREB protein can be found in a cell extract that has been pretreated with epinephrine or not, depending on your preference.Using an electrophoretic mobility shift experiment, you test the CREB protein's capacity to attach to a DNA fragment containing a cAMP response element (CRE). Specify what you hope to achieve.
(c) By binding one L-tryptophan molecule/monomer, the trp repressor binds to DNA to suppress syn-
thesis of L-tryptophan in E. coli. Below is the amino acid sequence of the helix – (reverse) turn – helix
region of the trp repressor that binds to DNA compared to the sequence of the corresponding DNA
binding motif of the Prl protein, a different type of repressor protein. A diagram of the trp repressor
dimer is also shown.
reverse turn
trp helix 4
70
Trp
-Gly-Glu-Met-Ser-Gln-Arg-Glu-Leu-Lys-Asn-Glu-Leu-Gly-Ala-Gly-
Ile-
Prl
-Ser-Glu-Glu-Ala-Lys-Glu-Glu-Leu-Ala-Lys-Lys-Cys-Gly-Ile-Thr-
Val-
Pri heilix
trp helix 5
80
90
Trp
Ala-Thr-Ile-Thr-Arg-Gly-Ser sgn-Ser-Leu-Lys-Ala-Ala-
Prl
Ser-Gln-Val-Ser-Asn-Trp-Phe-Gly-Asn-Lys-Arg-Ile-Arg-
Prl helix
Chapter 15 Solutions
Genetics: Analysis and Principles
Ch. 15.1 - 1. Combinatorial control refers to the phenomenon...Ch. 15.1 - 2. A regulatory transcription factor protein...Ch. 15.1 - 3. A bidirectional enhancer has the following...Ch. 15.1 - 4. Regulatory transcription factors can be...Ch. 15.2 - 1. A chromatin-remodeling complex may
a. change...Ch. 15.2 - Prob. 2COMQCh. 15.2 - 3. Which of the following characteristics is...Ch. 15.2 - 4. Transcriptional activation of eukaryotic genes...Ch. 15.3 - How can methylation affect transcription? a. It...Ch. 15.3 - 2. The process in which completely unmethylated...
Ch. 15.4 - Prob. 1COMQCh. 15.5 - The overall goal of the ENCODE Project is a. to...Ch. 15.6 - The binding of iron regulatory protein (IRP) to...Ch. 15 - Discuss the common points of control in eukaryotic...Ch. 15 - 2. Discuss the structure and function of...Ch. 15 - 3. What is meant by the term transcription factor...Ch. 15 - What are the functions of transcriptional...Ch. 15 - 5. Is each of the following statements true or...Ch. 15 - 6. Transcription factors usually contain one or...Ch. 15 - Prob. 7CONQCh. 15 - Prob. 8CONQCh. 15 - 9. Let’s suppose a mutation in the glucocorticoid...Ch. 15 - Prob. 10CONQCh. 15 - Prob. 11CONQCh. 15 - Prob. 12CONQCh. 15 - 13. Transcription factors such as the...Ch. 15 - An enhancer, located upstream from a gene, has the...Ch. 15 - 15. The DNA-binding domain of each CREB protein...Ch. 15 - The gene that encodes the enzyme called tyrosine...Ch. 15 - Prob. 17CONQCh. 15 - 18. What is a histone variant?
Ch. 15 - Prob. 19CONQCh. 15 - 20. What is meant by the term histone code? With...Ch. 15 - Prob. 21CONQCh. 15 - Histones are thought to be displaced as RNA...Ch. 15 - 23. What is an insulator? Describe two different...Ch. 15 - 24. What is DNA methylation? When we say that DNA...Ch. 15 - Lets suppose that a vertebrate organism carries a...Ch. 15 - 26. What is a CpG island? Where would you expect...Ch. 15 - Describe how the binding of iron regulatory...Ch. 15 - 1. Briefly describe the method of chromatin...Ch. 15 - Researchers can isolate a sample of cells, such as...Ch. 15 - Prob. 3EQCh. 15 - Prob. 4EQCh. 15 - Prob. 5EQCh. 15 - 6. As described in Chapter 21, an electrophoretic...Ch. 15 - Prob. 7EQCh. 15 - 1. Explain how DNA methylation could be used to...Ch. 15 - 2. Enhancers can occur almost anywhere in DNA and...
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