plasmid cloning vector
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What feature of a plasmid cloning vector facilitates selection for its presence in bacteria?
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- Ampicillin and tetracycline resistant genes are sometimes located on plasmids used in cloning. What is the importance of such genes in tranformation? What is the principle involved?Why do we need a plasmid to transform a bacteria? What are the essential components of a plasmid and explain their functions. What is the difference between a Plasmid and a Vector?During the colony selection at the end of the cloning step, there is a possibility of encountering false positive colonies (colonies containing the plasmid without the gene of interest). How does pZERO®-1 overcome this situation?
- How is a gene inserted into a plasmid cloning vector?What is the advantage that some plasmid cloning vectors carry the lacZ 'gene and the large number of cloning sites inserted into this gene?a) If lacZ was successfully inserted into the plasmid and E. coli successfully transformed, then the lacZ gene will be expressed which will breakdown X-gal producing a blue-colored colony. b) If the gene of interest was successfully inserted into the plasmid and E. coli successfully transformed, then the LacZ gene will not be expressed so X-gal will be broken down leading to a blue-colored colony. c) If the gene of interest was successfully inserted into the plasmid and E. coli successfully transformed, then the LacZ gene will not be expressed so X-gal will not be broken down leading to a white-colored colony. d) If lacZ was successfully inserted into the plasmid and E. coli successfully transformed, then the lacZ gene will be expressed which will breakdown X-gal producing a white-colored colony.
- Bacterial plasmids often serve as cloning vectors. Describe the essential features of a plasmid vector.What characteristics of plasmids and bacteriophages make themgood cloning vectors?What are the essential elements of a plasmid cloning vector? Discuss the importance of each element for dna cloning.
- When using a conventional plasmid cloning vector containing a b-galactosidase gene, it is possible to perform a "blue-white screen" to determine which bacteria have taken up a plasmid into which a DNA fragment as been inserted, as opposed to those that have taken up just reclosed plasmid vector, by growing the transformed cells on nutrient agar plates containing the artificial b-gal substrate X-gal. Will bacteria that have taken up a plasmid into which a DNA fragment has been inserted form a blue colony or a white colony when grown on this medium? Briefly explain why these bacteria would form a colony of the color you chose.What is a plasmid? Why is it an ideal tool in replicating genetic sequences?Why are antibiotic resistance genes usually included in plasmids used for genetic transformation?