Genetic Analysis: An Integrated Approach (3rd Edition)
3rd Edition
ISBN: 9780134605173
Author: Mark F. Sanders, John L. Bowman
Publisher: PEARSON
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Chapter 8, Problem 24P
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
a. List at least three 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?
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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?
There are similarities and differences during regulation of gene expression in both prokaryotes and eukaryotes. Promoters, transcription factors and RNA polymerase are essential elements in transcription but their properties and function may differ.a) Predict the outcome or consequences of mRNA transcription by RNA polymerase II in eukaryote without the presence of transcription factors (TF).
a. How do bacteria increase the efficiency of gene expression? Is this possible in eukaryotes?
b. A mutation in the promoter of Gene K disrupts an enzyme binding site and results in the loss of
Gene K expression. Is this change in gene expression likely happening at the transcriptional or the
translational level? Explain.
c. Propose three different mutations to prevent initiation, elongation, and termination of bacterial
transcription, respectively. Explain how/why each mutation would prevent its respective step. (Hint:
mutations can be in genes that encode proteins or regulatory DNA sequences)
Chapter 8 Solutions
Genetic Analysis: An Integrated Approach (3rd Edition)
Ch. 8 - Prob. 1PCh. 8 - 8.2 In one to two sentences each, describe the...Ch. 8 - 8.3 Answer these questions concerning...Ch. 8 - 8.4 The diagram below shows a DNA duplex. The...Ch. 8 - The following is a portion of an mRNA sequence:...Ch. 8 - Compare and contrast the properties of DNA...Ch. 8 - The DNA sequences shown below are from the...Ch. 8 - Bacterial and eukaryotic gene transcripts can...Ch. 8 - Describe the two types of transcription...Ch. 8 - What is the role of enhancer sequences in...
Ch. 8 - Prob. 11PCh. 8 - Draw a bacterial promoter and label its consensus...Ch. 8 - For a eukaryotic gene whose transcription require...Ch. 8 - Three genes identified in the diagram as A, B and...Ch. 8 - Prob. 15PCh. 8 - 8.16 The segment of the bacterial gene involved in...Ch. 8 - Prob. 17PCh. 8 - Prob. 18PCh. 8 - 8.19 A DNA fragment from the end of the mouse...Ch. 8 - 8.20 Wild-type E. coli grow best at but can grow...Ch. 8 - A mutant strain of Salmonella bacteria carries a...Ch. 8 - 8.22 The human wild-type allele and a certain...Ch. 8 - Prob. 23PCh. 8 - A full-length eukaryotic gene is inserted into a...Ch. 8 - The accompanying illustration shows a portion of a...Ch. 8 - DNA footprint protection (described in Research...Ch. 8 - Suppose you have a 1-kb segment of cloned DNA that...Ch. 8 - Assume that a mutation affects the gene for each...Ch. 8 - 8.29 The DNA sequence below gives the first base...Ch. 8 - 8.30 Genomic DNA from a mouse is isolated,...Ch. 8 - 8.31 A portion of a human gene is isolated from...
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- The IMD2 promoter contains three upstream transcription start sites (TSS) that are utilized under high GTP conditions and a single downstream TSS (-106) that is normally only utilized under low GTP conditions. In a wild type cell, expression of IMD2 mRNA only occurs if transcription initiates from the -106 TSS. In 300 words or less, describe: 1.) The normal function of Ssl2, and 2.) why a mutation in Ssl2, that increases its catalytic rate, would allow expression of the IMD2 ORF under high GTP conditions. (Conditions under which the IMD2 ORF is NOT expressed in the wild type.)arrow_forwardThe following diagram represents a transcription unit on a DNA molecule. a. Assume that this DNA molecule is from a bacterial cell. Draw the approximate locations of the promoter and terminator for this transcription unit. b. Assume that this DNA molecule is from a eukaryotic cell. Draw the approximate location of an RNA polymerase II promoter.arrow_forwardYou 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'arrow_forward
- Genes in both prokaryotes and eukaryotes are regulated by activators and repressors.a. Compare and contrast the mechanism of functionof a prokaryotic repressor (for example, Lac repressor) with a typical eukaryotic repressor protein(a direct repressor).b. Compare and contrast the mechanism of functionof a prokaryotic activator (for example, CAP) witha typical eukaryotic activator protein.arrow_forwardDescribe how transcription would be affected in the Galactose metabolizing pathway in Yeast in the presence of the following mutations. 1. A mutation that resulted in an inability of Gal80 to enter the nucleus. 2. A mutation that resulted in a lack of ability of Gal3 to bind galactose.arrow_forwardNegative supercoiling of DNA favors the transcription of genes because it facilitates unwinding. However, not all promoter sites are stimulated by negative supercoiling. The promoter site for topoisomerase II itself is a noteworthy exception. Negative supercoiling decreases the rate of transcription of this gene. Propose a possible mechanism for this effect and suggest a reason why it may occur.arrow_forward
- When the amino acid levels in eukaryotic cells are low, general protein synthesis is reduced. Gcn4 translation, however, is increased. A. What would happen under high and low amino acid conditions if only one of the upstream ORFs were deleted from Gcn4? B. What would happen under high and low amino acid conditions if all of the upstream ORFs were deleted from Gcn4?arrow_forwardTrinucleotide repeat expansions (TNREs) are associated with severaldifferent human inherited diseases. Certain types of TNREsproduce a long stretch of the amino acid glutamine within theencoded protein. When a TNRE exerts its detrimental effect byproducing a glutamine stretch, are the following statements true orfalse?A. The TNRE is within the coding sequence of the gene.B. The TNRE prevents RNA polymerase from transcribing thegene properly.C. The trinucleotide sequence is CAG.D. The trinucleotide sequence is CCG.arrow_forwardA full-length eukaryotic gene is inserted into a bacterial chromosome. The gene contains a complete promoter, polyadenylation sequence and a wildtype nucleotide sequence. However, the gene does not produce a functional protein List four reasons why this gene is not expressed in bacteria List five changes you recommend that allow expression of this eukaryotic gene in bacteriaarrow_forward
- The MRNA for a mammalian peptide that enhances electrolyte transfer to reduce paralysis was spliced then reversed transcribed and the CDNA ligated into a phage and transfected into host bacteria cells. The host then transcribed and translated this. The protein product however was non-functional. What is the most probable reason for this result? A. Absence of post-transcriptional modification in bacterial cells B. Differences in the post-translational mechanisms in eukaryotes and prokaryotes. C. Differences in the translational mechanisms involved in prokaryotes, e.g. 70 S ribosome vs 80S ribosome. D. The mRNA did not contain a promoter region to allow peptide translation. E. Insertion of portions of the phage DNA which added sequences for amino acids changing the function of the protein product.arrow_forwardAnother class of suppressor mutations, not describedin the chapter, are mutations that suppress missensemutations.a. Why would bacterial strains carrying such missense suppressor mutations generally grow moreslowly than strains carrying nonsense suppressormutations?arrow_forwardDiscuss how the expression of a protein can be regulated post transcription in eukaryotic cells through, using the following key terms: Degradation of mRNA (two ways) Blocking translation Degradation of the proteinarrow_forward
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