Microbiology: An Evolving Science (Fourth Edition)
4th Edition
ISBN: 9780393615098
Author: John W. Foster, Joan L. Slonczewski
Publisher: W. W. Norton & Company
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Chapter 12, Problem 7RQ
Summary Introduction
To review:
The important considerations for the construction of a plasmid synthesizing His6 (histidine)-tagged protein.
Introduction:
A circular double-stranded DNA (deoxyribonucleic acid) molecule that replicates independently is known as a plasmid. Plasmids exist naturally in bacterial cells to provide special benefits like antibiotic resistance. His6-tagged proteins can be identified by the presence of a string of histidine residues that can bind to nickel under suitable environment.
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Chapter 12 Solutions
Microbiology: An Evolving Science (Fourth Edition)
Ch. 12.1 - Prob. 1TQCh. 12.1 - Prob. 2TQCh. 12.1 - Prob. 3TQCh. 12.1 - Prob. 4TQCh. 12.2 - Prob. 1TQCh. 12.2 - Prob. 2TQCh. 12.2 - Prob. 3TQCh. 12.3 - Prob. 1TQCh. 12.4 - Prob. 1TQCh. 12.4 - Prob. 2TQ
Ch. 12.5 - Prob. 1TQCh. 12 - Prob. 1RQCh. 12 - Prob. 2RQCh. 12 - Prob. 3RQCh. 12 - Prob. 4RQCh. 12 - Prob. 5RQCh. 12 - Prob. 6RQCh. 12 - Prob. 7RQCh. 12 - Prob. 8RQCh. 12 - Prob. 9RQCh. 12 - Prob. 10RQCh. 12 - Prob. 11RQCh. 12 - Prob. 12RQCh. 12 - Prob. 13RQCh. 12 - Prob. 14RQCh. 12 - Prob. 15RQCh. 12 - Prob. 1TQCh. 12 - Prob. 2TQCh. 12 - Prob. 3TQ
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- A researcher has a Trp− auxotrophic strain of E. coliwith a mutation in a single gene. To identify thatmutant gene, she uses a genomic library made from a wild-type version of that same strain to find plasmids that rescue the mutant phenotype. The result issurprising. She recovers 10 plasmids that provide aTrp+ phenotype, but six of the plasmids contain geneX, while the other four contain gene Y. Our scientisthas encountered a phenomenon called multicopysuppression, related to the fact that plasmids are usually present in several copies per bacterium. Becausethe genes in the plasmids are present in more thantheir usual single copy in the bacterial chromosome,more than the usual amount of Protein X or Protein Yis being produced from the plasmids. Sometimes,overexpression of one protein can rescue the mutantphenotype caused by loss of a different protein.Suggest at least two ways that our scientist could determine which of the two genes, gene X or gene Y, actually corresponds to the…arrow_forwardWhat is the role of ‘Ori’ in any plasmid?arrow_forwardYou first need to create a plasmid. What are the minimal components necessary to develop this plasmid? In addition to these components, please draw the plasmid. In this illustration, I am looking for the components, the direction of transcription (i.e., the direction the genes will be transcribed), and what should be transcribed last? Also, where would you specifically insert the P. falciparum gene in this plasmid, and why are you checking reading frames in your gene? Finally, please name your plasmid using the correct nomenclature too. You are excited you have this new plasmid; you want to transfect it into P. marinus and provide it as an oral vaccine to laboratory mice. However, even though your supervisor enjoys your enthusiastic attitude, they do not allow you to do this quite yet because all you have is this plasmid. Why doesn’t your supervisor allow you to use the laboratory mouse for this research regarding animal welfare guidelines? Your answer should include the 3 R’s of…arrow_forward
- Explain the basis for the following statement: Transcription of two genes on a plasmid can occur without the concomitant action of a topoisomerase, but only if those two genes are oriented in opposite directions.arrow_forwardWhat is the direction of transcription in the plasmid shown below? Explain your answer based on the features present in the figure.arrow_forwardIn the genotype presented (genomic plus plasmid genes), which of the following statements Is phenotype? genome: IS P O`z+Y° plasmid: I* Pt otzY+ no production of either B-gal and permease O B-gal and permease induced when lactose is present no B-gal is produced and expression of permease when lactose is present constitutive expression of B-gal and permease constitutive expression of B-gal and no expression of permeasearrow_forward
- Examine the structure of the pBR322 plasmid depicted below. Assume total size of the plasmid is 4,361 bp and the blue numbers indicate locations of restriction sites relative to the O point at the top of the plasmid. What size fragments would be generated by the following restriction digestion reactions? 1. Sall 2. Sal 1 + BamH1 3. Sal 1 + EcoR1 4. Sal I + BamH1 + EcoR1 PstI 3607 3000 4000 amp ori HindIII Edit View Insert Format Tools Table EcoRI EcoRV 4359 0 29 185 pBR322 4361 bp 2295 NdeI tet 2000 BamHI 375 651 SalI 1000 Type a short answer in the space provided below.arrow_forwardWhat do you mean by vector plasmid?arrow_forwardWhen plasmids are isolated from bacterial cells, they may existin a number of forms.a. List the different forms that may be found.b. Which do you think would migrate the fastest and farthest in anelectrophoresis experiment and why?arrow_forward
- The transfection reagent used to introduce plasmids to the HEK293 cells was a lipid reagent. Why is lipid utilized for this procedure?arrow_forwardBesides plasmid, what other vectors could you use to amplify your gene of interest? Name THREE (3) other vectorsarrow_forwardYou have set up a recombinant DNA experiment using the plasmid PBR322 as the vector (see plasmid below). You use the BamHI restriction site on the plasmid to insert the target DNA. The plasmid is then used to transform E.coli colls Is the following statement True or False? Growth of the transformed cells on agar containing both ampicillin and tetracycline will eliminate any cells that do not contain a plasmid. Clal Hindlll EcoRI Pvul BamHI Pstl amp tet PBR322 -Sall ori rop Pvull True Falsearrow_forward
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