Ocean pout, a fish found in co
Ocean pout, a fish found in cold waters, produce anantifreeze protein that allows them to survive at near freezingtemperatures. Your company plans to genetically engineer bacteriato grow antifreeze proteins to improve the storage of ice creamproducts
a) The vector and PCR product that you will use toclone the antifreeze gene are shown above along with the locationsof three different restriction enzyme cut sites Which restrictionenzyme (Hindill, BamH1, or EcoR1) will allow you to successfullyclone this transgeno? Brietly justify your answer.
b) Describe how you will select for bacteria thatcarry the recombinant DNA molecule once you have completed thetransformation
Answer:
A) BamH1 restriction enzyme is used to cut the vector and geneof interest. This will allow succesful cloning of the gene. Becausethe BamH1 gene is found outside the coding region of gene in geneof interest. And it is also present in ampar region in the plasmid.This further helps in selection of recombinant bacteria. B) You canput both kind of bacteria in presence of UV light. You will findthat recombinant bacteria will glow in presence of UV because ofGFP protein. But the non recombinant bacteria will not glow. Inthis way, you can isolate the recombinant bacteria. other method,but not so much useful: The bacteria is grown in presence ofampicilin containing media. The bacteria which has recombinantplasmid will not grow in presence of ampicillin containing mediabecause ampar gene is non functional in recombinant bacteriabecause of insertional inactivation. Where as other untransformedbacteria will grow. So,first method is best.
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