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 9P
Figure 18.6 shows a picture of Glofish ®, transgenic zebrafish (Danio rerio) that express GFP, RFP, and some of their derivatives. Glofish® were the first GM pets. Genes can be transferred into the zebrafish genome using a transposable element called Tol2 from medaka fish (Oryzias latipes). The Tol2 transposon serves as a vector for gene transfer in a manner similar to Drosophila P elements as shown in Fig. 18.4. To use the Tol2 vector, researchers inject two-cell stage zebrafish embryos with two DNAs: One DNA expresses the Tol2 transposase gene, while the other recombinant transposon contains the transgene. If the transposon hops into a chromosome of a germ-line cell, a stable line of transgenic fish can be established.
Scientists have synthesized several different genes encoding derivatives of GFP and RFP. The derivatives have amino acid changes that alter their color, so that fish glowing green, red, orange, blue, or purple are available in many pet stores.
| a. | Diagram the two recombinant DNA constructs needed to generate the fluorescent fish. |
| b | Why is a transposon from medaka fish used rather than a zebrafish transposon? |
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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 uses the term homologous correctly?A. The two X chromosomes in female mammalian cells are homologousto each other.B. The α-tubulin gene in Saccharomyces cerevisiae is homologousto the α-tubulin gene in Arabidopsis thaliana.C. The promoter of the lac operon is homologous to the promoterof the trp operon.D. The lacY genes of E. coli and K. pneumoniae are approximately60% homologous to each other.
i) Suppose we want to insert the GFP sequence after the promoter of a gene X to create a fusion protein. Please describe the strategy that you will follow to edit this gene with CRISPR-Cas9 technology, in order to visualize the expression of gene X in live cells.
(ii) Please explain how a knockout mouse for a gene X can be produced with CRISPR-Cas9 technology
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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