Genetics: From Genes to Genomes
6th Edition
ISBN: 9781259700903
Author: Leland Hartwell Dr., Michael L. Goldberg Professor Dr., Janice Fischer, Leroy Hood Dr.
Publisher: McGraw-Hill Education
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Chapter 18, Problem 20P
One way to determine where inside a cell a protein (protein X) normally localizes is to generate a reporter gene construct containing: (i) the gene X regulatory region and coding sequences, and (ii) coding sequences for GFP fused in frame to the 3′ end of the gene X coding sequences just before the stop codon. A mouse containing such a transgene will express a hybrid protein X-GFP only in those cells in which gene X is normally expressed.
| a. | The gene X-GFP fusion gene described could be generated by knocking in GFP coding sequences instead of by random insertion of a transgene. Diagram the knockin construct you could use for this purpose. |
| b. | What might the advantage be of the knockin strategy versus the transgene strategy? |
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. One way to determine where inside a cell a protein(protein X) normally localizes is to generate a reportergene construct containing: (i) the gene X regulatoryregion and coding sequences, and (ii) coding sequencesfor GFP fused in frame to the 3′ end of the gene Xcoding sequences just before the stop codon. A mousecontaining such a transgene will express a hybrid protein X-GFP only in those cells in which gene X is normally expressed.a. The gene X-GFP fusion gene described could begenerated by knocking in GFP coding sequencesinstead of by random insertion of a transgene.Diagram the knockin construct you could use forthis purpose.b. What might the advantage be of the knockin strategy versus the transgene strategy?
(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
You would like to add a nuclear localization sequence (NLS) of Lys-Lys-Lys-Arg-Lys to a
protein that is usually found in the cytoplasm of a yeast cell. To accomplish this, you
introduce the nucleotide sequence encoding the NLS into the gene that encodes the
cytoplasmic protein of interest.
a. What is the size of the nucleotide insert that will encode the NLS? Briefly explain.
5'
3'
b. Below is a diagram of the gene encoding the cytoplasmic protein of interest in
the yeast genome. If your goal is to put the NLS at the carboxyl (C) terminus of
the protein, at which location (A-E) should the NLS be inserted? Briefly explain.
A
TATAA
ATATT
promoter
+1
B
ATG
TAC
D
TAA
ATT
stop
codon
E
3'
5'
Chapter 18 Solutions
Genetics: From Genes to Genomes
Ch. 18 - Match each of the terms in the left column to the...Ch. 18 - Mice are usually gray, but a mouse geneticist has...Ch. 18 - Sometimes, genes transferred into the mouse genome...Ch. 18 - In mice, a group of so-called Hox genes encode...Ch. 18 - The fly eyes shown in Fig. 18.7 are malformed...Ch. 18 - This problem concerns a technique called enhancer...Ch. 18 - Fish and other organisms that live in the Arctic...Ch. 18 - a. Describe two ways you could potentially make a...Ch. 18 - Figure 18.6 shows a picture of Glofish ,...Ch. 18 - Some people are concerned about the possible...
Ch. 18 - The goal of the Knockout Mouse Project is to...Ch. 18 - Prob. 12PCh. 18 - Prob. 13PCh. 18 - a. Which genome manipulation technique would you...Ch. 18 - a. Diagram a knockin construct that could have...Ch. 18 - Prob. 16PCh. 18 - Prob. 17PCh. 18 - The transcription factor Pax6 is required...Ch. 18 - Mouse models for human genetic diseases are...Ch. 18 - One way to determine where inside a cell a protein...Ch. 18 - In Problem 5 in Chapter 17, you saw that a SNP...Ch. 18 - Scientists now routinely use CRISPR/Cas9 to make...Ch. 18 - Geneticists are currently considering using...Ch. 18 - a. Figures 18.9 and 18.12 demonstrated methods to...Ch. 18 - Nonhomologous end-joining NHEJ of a double-strand...Ch. 18 - One problem that researchers sometimes encounter...Ch. 18 - Researchers at the University of California at San...Ch. 18 - Prob. 28PCh. 18 - F. Port and S. Bullock at the University of...Ch. 18 - On Fig 18.14, locate the PAM site and identify the...Ch. 18 - Prob. 31PCh. 18 - Prob. 32PCh. 18 - Recall that Leber congenital amaurosis LCA, a form...Ch. 18 - One potential strategy for gene therapy to correct...Ch. 18 - Recently, scientists have used a mouse model for...
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