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 7.6, Problem 2TQ
Summary Introduction
To review:
The possibility of transferring a foreign gene into the bacteria which possess endonuclease restriction enzyme.
Introduction:
The enzyme that breaks the DNA (deoxyribonucleic acid) into segments by making cuts at specific restriction or recognition sites is the restriction enzyme. These enzymes are also known as endonuclease enzymes that break the phosphodiester bond in the DNA molecules.
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Which of the following is necessary for a PCR reaction to proceed? a) the sequence of the ends of the DNA to be amplified must be known. b) the sequence of restriction endonuclease recognition sites in the DNA to be amplified and in the plasmid, where the amplified DNA fragment will be cloned must be known. c) The complete sequence of the DNA to be amplified must be known. d) The sequence of restriction endonuclease recognition sites in the DNA to be amplified must be known
What sequence do the restriction enzymes used in the lab recognize? How do they cut? And how would these different cuts effect cloning? (i.e. compare and contrast how overhang and blunt cuts will differ with cloning.)
What organisms were the restriction enzymes used in this lab derived from?
Why are restriction enzymes important for the organism that makes them?
What is the purpose of multiple cloning sites on pUC19? What size were each of your plasmid fragments?
If you use the pUC18 vector to clone in the MCS region, predict the following:
a) Do bacteria that are blue in color have a cloned insert?
b)Do bacteria that are white in color have a cloned insert?
c) If you were to grow these cells on Chloramphenicol (an antibiotic), would the bacteria with the pUC plasmid grow? Why or Why not?
Chapter 7 Solutions
Microbiology: An Evolving Science (Fourth Edition)
Ch. 7.2 - Prob. 1TQCh. 7.2 - Prob. 2TQCh. 7.2 - Prob. 3TQCh. 7.2 - Prob. 4TQCh. 7.3 - Prob. 1TQCh. 7.3 - Prob. 2TQCh. 7.3 - Prob. 3TQCh. 7.3 - Prob. 4TQCh. 7.3 - Prob. 5TQCh. 7.6 - Prob. 1TQ
Ch. 7.6 - Prob. 2TQCh. 7.6 - Prob. 3TQCh. 7 - Prob. 1RQCh. 7 - Prob. 2RQCh. 7 - Prob. 3RQCh. 7 - Prob. 4RQCh. 7 - Prob. 5RQCh. 7 - Prob. 6RQCh. 7 - Prob. 7RQCh. 7 - Prob. 8RQCh. 7 - Prob. 9RQCh. 7 - Prob. 10RQCh. 7 - Prob. 11RQCh. 7 - Prob. 12RQCh. 7 - Prob. 13RQCh. 7 - Prob. 14RQCh. 7 - Prob. 15RQCh. 7 - Prob. 1TQCh. 7 - Prob. 2TQCh. 7 - Prob. 3TQ
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- Which of the following can be termed as a restriction modification system?a) Restriction endonuclease + methylaseb) DNA ligase + methylasec) Restriction endonuclease + acetylased) DNA ligase + acetylasearrow_forwardWhy are nucleic acid probes necessary to lyse cells containing plasmids in order to detect the product of the cloned gene?arrow_forwardDescribe two features that are characteristic of the action of restriction endonucleases. How do these features make restriction endonucleases useful to genetic engineers?arrow_forward
- Why do bacteria make restriction endonucleases? What is it about the endonucleases that prevents bacteria from destroying their own DNA?arrow_forwardWhy do restriction endonucleases not hydrolyze DNA from the organism that produces it?arrow_forwardWhy do bacteria make restriction endonucleases?arrow_forward
- Restriction sites are palindromic; that is, they read the same in the5' to 3' direction on each strand of DNA. What is the advantage ofhaving restriction sites organized this way?arrow_forwardIn the formation of recombinant DNA, a restriction endonuclease cuts a bacterial plasmid to give sticky ends. The DNA segments that are to be added to the plasmid are cleaved with the same restriction endonuclease. What aresticky ends and why is it important that the target DNA and the plasmid it will be incorporated into have complementary sticky ends?arrow_forwardBacteriophage lambda (λ) consists primarily of a head, which contains the genomic DNA, and a tail that is involved in phage attachment to bacterial cells. The diagram to the right represents the bacteriophage lambda genomic DNA, showing the locations of important gene clusters (Ausubel et al. 1998). Arrows below mark the sites where the restriction enzyme HindIII cuts the DNA, and the numbers indicate the number of base pairs in each fragment. The restriction enzyme HindIII cuts the λ genome 7 times, how many fragments are produced following this reaction?arrow_forward
- We have two specific strains of E. coli that have shown horizontal gene transfer (HGT) when mixed. To experimentally determine the method of HGT that is happening, the following conditions are set up in different tubes of culture media: A) Donor and recipient strain mixed together (control - no treatment). B) Donor and recipient strains mixed together, DNase added (can digest DNA in solution, not within cells).C) Special tube containing a membrane filter (with pores that allow DNA and viruses to pass through, but not bacterial cells) that separates two compartments. Donor strain is added on one side, the recipient strain on the other (they are separated by the filter).D) Donor and recipient strains mixed together, with chemical that inactivates viruses (chemical affects bacteriophages in solution so they are unable to attach to cells). The results: Tubes A, B, and D: HGT was observed. Tube C: HGT was NOT observed. Based on this, which type of HGT was occurring? Conjugation,…arrow_forwarda）What two restriction sites are you going to use to clone your PCR product into the pL4440 plasmid? What are their DNA sequence? b) State the primer sequence you will use to amplify the F27C1.7 gene ready to be cloned into the pL4440 plasmid? c) How would you go about cloning this amplified DNA into pL4440? Using your knowledge of cloning list 5 important aspects of the method.arrow_forwardA 12 kb linear DNA fragment is subject to single or double RE digest and agarose gelelectrophoresis, to yield the gel profile shown below. The first lane contains the size marker(M).a) Explain how the name of the enzyme EcoRI is derived.b) How many sites are there for EcoRI and PvuII respectively on this DNA fragment?c) Use the sizes of the DNA bands on the gel to compile a restriction enzyme map of the DNAfragment. Indicate the positions of the restriction enzymes sites for EcoRI and PvuII on themap.arrow_forward
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