GENETIC ANALYSIS: AN INTEG. APP. W/MAS
2nd Edition
ISBN: 9781323142790
Author: Sanders
Publisher: Pearson Custom Publishing
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Chapter 16, Problem 9P
Translational fusions between a protein of interest and a reporter protein are used to determine the subcellular location of proteins in vivo. However, fusion to a reporter protein sometimes renders the protein of interest non-functional because the addition of the reporter protein interferes with proper protein folding, enzymatic activity, or protein
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A full-length eukaryotic gene is inserted into a bacterial chromosome. The gene contains a complete promoter sequence and a functional polyadenylation sequence, and it has wild-type nucleotides throughout the transcribed region. However, the gene fails to produce a functional protein.
a)List at least 3 possible reasons why this eukaryotic gene is not expressed in bacteria.
b)What changes would you recommend to permit expression of this eukaryotic gene in a bacterial cell?
Give typing answer with explanation and conclusion
a) List three eukaryotic gene expression mechanisms that do not occur in prokaryotes. For two of these, give specific examples and the functional outcomes.
b) Describe what is meant by the term “RNA silencing”.
c) Using diagrams, give two examples of RNA silencing mechanisms and indicate one difference.
In the absence of tryptophan and the trpD structural gene contains a frameshift insertion mutation, the structural genes will: [ Select ] ["No longer be expressed", "Be constantly expressed but the protein will be truncated and possibly non-functional", "Be constantly expressed but the protein will be longer and possibly non-functional"]
In the presence of tryptophan and the trpD structural gene contains a frameshift insertion mutation, the structural genes will: [ Select ] ["No longer be expressed", "Be constantly expressed but the protein will be truncated and possibly non-functional", "Be constantly expressed but the protein will be longer and possibly non-functional"]
Chapter 16 Solutions
GENETIC ANALYSIS: AN INTEG. APP. W/MAS
Ch. 16 - 14.1 What are the advantages and disadvantages of...Ch. 16 - Prob. 2PCh. 16 - 3. Genetic maps and physical maps are both...Ch. 16 - 14.5 What are the advantages and disadvantages of...Ch. 16 - 14.6 You have cloned the mouse ortholog (see...Ch. 16 - 14.13 The CBF genes of Arabidopsis are induced by...Ch. 16 - 14.14 When the S. cerevisiae genome was sequenced,...Ch. 16 - 14.15 Translational fusions between a protein of...Ch. 16 - In enhancer trapping experiments, a minimal...Ch. 16 - 14.19 In Genetic Analysis, we designed a screen to...
Ch. 16 - How would you design a genetic screen to find...Ch. 16 - 14.21 The eyes of Drosophila develop from imaginal...Ch. 16 - 14.22 Given your knowledge of the genetic tools...Ch. 16 - Mutations in the CFTR gene result in cystic...Ch. 16 - Prob. 16PCh. 16 - 14.25 How would you conduct a screen to identify...Ch. 16 - In land plants, there is an alternation of...Ch. 16 - 14.27 The Drosophila evenskipped (eve) gene is...Ch. 16 - Prob. 20PCh. 16 - 14.29 As shown in Figure, mutations in the...
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- A series of exonuclease deletions were used to study the promoter of the rice hemA gene, giving the results shown below (the intact promoter is on top). Based on these results, what conclusions could you draw from each of the deletion construct, and what do you know about the nature of the promoter? Relative activity -500 Reporter gene 100% Reporter gene 180% -350 -250- Reporter gene 180% 75% -150 Reporter gene 45% Reporter gene -100 0% - 8 Reporter genearrow_forwardA number of foreign proteins have been expressed in bacterial and mammalian cells. Describe the essential features of a recombinant plasmid that are required for the expression of a foreign gene. How can the foreign protein be modified to facilitate its purification? What is the advantage of expressing a protein in mammalian cells versus bacteria?arrow_forwardPROTEIN X HAS THE POTENTIAL FOR MEDICAL APPLICATION IN HUMAN. IT IS ORIGINALLY FROM A PLANT AND HAD BEEN SUCCESSFULLY CLONED FROM ITS GENE. AFTER MANY ATTEMPTS TO CLONE IT INTO EUKARYOTIC EXPRESSION VECTORS WERE UNSUCCESSFUL, THE RESEARCHER DECIDED TO USE E.COLI EXPRESSION VECTOR. HOW THE RECOMBINANT PROTEIN COULD BE EXPRESSED IN E.COLI EXPRESSION VECTOR.1. FACTORS AFFECTING BACTERIAL EXPRESSION SYSTEM.2. STRATEGIES TO IMPROVE THE PROTEIN EXPRESSION.3. WAYS TO ENSURE THE PROTEIN IS ACTIVELY PRODUCED.arrow_forward
- When a region of DNA that contains the genetic information for a protein is isolated from a bacterial cell and inserted into a eukaryotic cell in a proper position between a promoter and a terminator, the resulting cell usually produces the correct protein. But when the experiment is done in the reverse direction (eukaryotic DNA into a bacterial cell), the correct protein is often not produced. Can you suggest an explanation?arrow_forwardWhen Laybourne and Kadonaga studied the effects of histone proteins on eukaryotic transcription using an in vitro transcription assay explain why: a) they used two different DNA templates that contained different promoter structures. b) when they included both activator protein and histones, they always added the histone proteins before adding the activator to the transcription assay mixture. (Ctri) -arrow_forwardWhich of the following statements is true regarding the lys2-128d reporter? Select all that apply. a.) Mutants that activate this reporter are likely to have mutations in PIC components. b.) This reporter is sensitive to changes in chromatin maintenance because the start site is located within the ORF. c.) Mutants that activate this reporter cause downstream shifts in transcription start site selection. d.) This reporter is being used to help us rule out mutations that are likely impacting chromatin structure.arrow_forward
- Consider the following simple regulatory pathways. Assume the full pathway is shown. A- E- B- F- C- G- D- 1 H- A You identify several null mutations (a complete deletion of the gene). For each mutant (ind with a - sign), determine whether the final product (I, J, K or L) is inducible, uninducible, or constitutive. 2 B 3 C 4 D inducible inducible constitutive uninducible constitutive inducible inducible E uninducible F G H > > >arrow_forwardPROTEIN X HAS THE POTENTIAL FOR MEDICAL APPLICATION IN HUMAN. IT IS ORIGINALLY FROM A PLANT AND HAD BEEN SUCCESSFULLY CLONED FROM ITS GENE. AFTER MANY ATTEMPTS TO CLONE IT INTO EUKARYOTIC EXPRESSION VECTORS WERE UNSUCCESSFUL, THE RESEARCHER DECIDED TO USE E.COLI EXPRESSION VECTOR. HOW THE RECOMBINANT PROTEIN COULD BE EXPRESSED IN E.COLI EXPRESSION VECTOR.1. STRATEGIES TO IMPROVE THE PROTEIN EXPRESSION.arrow_forwardResearchers have identified a gene (FR) responsible for watermelon resistance to infection by Dacus curcurbitae (a close relative of Drosophila melanogaster). They isolate RNA from resistant (FR+) and sensitive (fr-) watermelons and use a probe that will recognize both FR+ and fr- transcripts. They also isolate protein from resistant and sensitive watermelons and perform a Western blot using an antibody that can recognize the fr- and FR+ protein. Describe the results illustrated below and give a plausible molecular explanation for these observations.arrow_forward
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