Microbiology With Diseases By Taxonomy (6th Edition)
6th Edition
ISBN: 9780134832302
Author: Robert W. Bauman Ph.D.
Publisher: PEARSON
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Chapter 8, Problem 9CT
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A shuttle vector is a vector (usually a plasmid) constructed so that it can propagate in two
different host species. One of the most common types of shuttle vectors is the
yeast shuttle
vector. Examples of such vectors derived from yeast are Yeast Episomal Plasmid (YEP),
Yeast Integrating Plasmid (YIP) and Yeast Replicating Plasmid (YRP). Why is YEP preferred
over YIP and YRP? Give your thoughts on this.
A full-length eukaryotic gene is inserted into a bacterial chromosome. The gene contains a complete promoter, a wildtype nucleotide sequence and a polyadenylation sequence. However, the gene does not produce a functional protein.
Students may submit their answers as bullet points or a table to facilitate grading.
Part A: List up to five reasons why this gene is not expressed in E. coli.
Part B: What changes do you recommend that would facilitate expression of the eukaryotic gene in E. coli.
The entire genome of the yeast Saccharomyces cerevisiae has been sequenced. This sequencing has led to the identification of all the open reading frames (ORFs, gene-size sequences with appropriate translational initiation and termination signals) in the genome. Some of these ORFs are previously known genes with established functions; however, the remainder are unassigned reading frames (URFs). To deduce the possible functions of the URFs, they are being systematically, one at a time, converted into null alleles by in vitro knockout techniques. The results are as follows:15 percent are lethal when knocked out.25 percent show some mutant phenotype (altered morphology, altered nutrition, and so forth).60 percent show no detectable mutant phenotype at all and resemble wild type.Explain the possible molecular-genetic basis of these three mutant categories, inventing examples where possible.
Chapter 8 Solutions
Microbiology With Diseases By Taxonomy (6th Edition)
Ch. 8 - Why did the discovery and development of...Ch. 8 - Why wasnt polymerase chain reaction (PCR)...Ch. 8 - Why dont doctors routinely insert genes into their...Ch. 8 - Why dont scientists who work with recombinant DNA...Ch. 8 - Which of the following statements is true...Ch. 8 - A DNA gene synthesized from an RNA template is...Ch. 8 - Prob. 3MCCh. 8 - Prob. 4MCCh. 8 - Prob. 5MCCh. 8 - Prob. 7MC
Ch. 8 - Prob. 9MCCh. 8 - Prob. 10MCCh. 8 - Prob. 1MTFCh. 8 - Prob. 2MTFCh. 8 - Prob. 3MTFCh. 8 - ________ Protoplast fusion is often used in the...Ch. 8 - Prob. 5MTFCh. 8 - Describe three artificial methods of introducing...Ch. 8 - Prob. 2SACh. 8 - Prob. 3SACh. 8 - List three potential problems of recombinant DNA...Ch. 8 - Examine the restriction sites listed in Table 8.1...Ch. 8 - Prob. 2CTCh. 8 - A thermocycler uses DNA polymerase from...Ch. 8 - How is the result of a Southern blot similar to...Ch. 8 - Prob. 6CTCh. 8 - Prob. 8CTCh. 8 - Prob. 9CTCh. 8 - Using the following terms, fill in the following...
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- Here is a eukaryotic gene. The numbers given are base pairs of exon and intron. How long in bases will the pre mRNA transcript be? Explain briefly. What is the maximum number of amino acids that could make up the protein product from the final mRNA? Explain briefly.arrow_forwardThe technique of fluorescence in situ hybridization (FISH) is described. This is another method for examining sequence complexity within a genome. In this method, a DNA sequence, such as a particular gene sequence, can be detected within an intact chromosome by using a DNA probe that is complementary to the sequence.For example, let’s consider the β-globin gene, which isfound on human chromosome 11. A probe complementary to theβ-globin gene binds to that gene and shows up as a brightly colored spot on human chromosome 11. In this way, researchers can detectwhere the β-globin gene is located within a set of chromosomes. Becausethe β-globin gene is unique and because human cells are diploid(i.e., have two copies of each chromosome), a FISH experimentshows two bright spots per cell; the probe binds to each copy ofchromosome 11. What would you expect to see if you used thefollowing types of probes?A. A probe complementary to the Alu sequenceB. A probe complementary to a tandem array near…arrow_forwardThe DNA sequence of one strand of a gene from threeindependently isolated mutants is given here (5′ endsare at left). Using this information, what is the sequence of the wild-type gene in this region?mutant 1 ACCGTAATCGACTGGTAAACTTTGCGCGmutant 2 ACCGTAGTCGACCGGTAAACTTTGCGCGmutant 3 ACCGTAGTCGACTGGTTAACTTTGCGCGarrow_forward
- Ihsan is a biologist working with the genetics of a psychrophilic bacterium. He cloned an antifreeze gene from the bacteria for further analysis. After cloning, he isolated the plasmid carrying his gene of interest for sequencing. Ihsan finally received the nucleotide sequence of his gene. Explain in detail how he could verify whether the nucleotide sequence matches his gene of interest using the bioinformatics databases available.arrow_forwardWhen cloning a foreign DNA fragment into a plasmid, it is often useful to insert the fragment at a site that interrupts a selectable marker(such as the tetracycline-resistance gene of pBR322). The loss of function of the interrupted gene can be used to identify clones containing recombinant plasmids with foreign DNA. With a yeast artificial chromosome (YAC) vector, it is not necessary to do this; the researcher can still distinguish vectors that incorporate large foreign DNA fragments from those that do not. How are these recombinant vectors identified?arrow_forwardA shuttle vector is a vector (usually a plasmid) constructed so that it can propagate in two different host species. One of the most common types of shuttle vectors is the yeast shuttle vector. Examples of such vectors derived from yeast are Yeast Episomal Plasmid (YEp), Yeast Integrating Plasmid (YIp) and Yeast Replicating Plasmid (YRp). Among these three vectors, YIp has the lowest transformation frequency and copy number per cell. Explain why Ylp is still popularly used despite its limitations.arrow_forward
- Shown below is a diagram for a plasmid vector you want to use to clone a gene. The diagram shows the location of the recognition sites for four restrictions enzymes, BamHI (B), EdoRI (E), Hindill (H), and Xhol (X). The genes encoding beta-lactamase (AmpR) and beta-galactosidase (lacZ) are indicated. If you were to use this vector, which enzyme should be used to linearize the plasmid in preparation for cloning? E B lacz O Hindi!! BamHI O EcoRI O Xhol H EcoRI and Xhol E -X AmpRarrow_forwardA scientist seeks to synthesize a rare human protein in lab. To accomplish this, she utilizes recombinant techniques to insert the DNA of a eukaryotic gene for the protein into bacterial plasmids. These plasmids are transformed into bacteria for expression. She is disappointed to discover that the gene product from the bacteria is not the correct protein. What corrective step could she take in the procedure to fix this issue?arrow_forwardA biologist is attempting to clone the gene encoding a particular enzyme (Enz) into a plasmid vector in E.coli. This plasmid has a gene encoding a green fluorescent protein (GFP) as well as a gene for tetracycline antibiotic resistance (TetR). The restriction site (to clone foreign DNA into) is within the GFP sequence. Which of the following would be expected when trying to see which E. coli cells acquired the recombinant plasmid (i.e., carrying the Enz gene)? Bacteria UNABLE to grow on tetracycline-containing media AND are NOT able to make green fluorescent protein are the ones that contain the recombinant plasmid. Bacteria able to grow on tetracycline-containing media AND that are NOT able to make green fluorescent protein are the ones that contain the recombinant plasmid. Bacteria able to grow on tetracycline-containing media AND are able to make green fluorescent protein are the ones that contain the recombinant plasmid. Bacteria UNABLE to…arrow_forward
- Mouse genomic DNA is treated with a restriction endonuclease and electrophoresed in an agarose gel. A radioactive probe made from the human gene rxr-1 is used to perform a Southern blot. The experiment was repeated three times. Explain the results of these repeated experiments:arrow_forwardDescribe how you would use replica plating of mutagenized, haploid yeast cells to identify temperature-sensitive (ts) mutations in essential genes needed for yeast growth and survival.arrow_forwardYou are studying a gene containing three exons (exon 1 = 66 nucleotides , exon 2 = 99 nucleotides , and exon 3 = 333 nucleotides ) that produces a primary RNA transcript 748 nucleotides long . Gel electrophoresis analysis shows different size mature transcripts are produced in three different cell types . The sizes of the mature transcripts are the following : 1. Neuronal nalcells = 498 nucleotides 2. Muscle cells = 600 nucleotides 3. Liver cells = 399nu nucleotides Primary RNA ranscript = 748 nucleotides a. Name the strategy that allows the same gene to produce different mature transcripts in different tissues . b. Describe the ways (or combinations of exons /introns ) used to produce each one ofthe three mature transcripts . Label the each mature transcripts and include its length . c. Indicate the length of the polypeptide encoded by each mature transcriptarrow_forward
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