To create a recombinant molecule, restriction enzymes are used to generate fragments of both vector and insert DNA; DNA ligase is then used to join those fragments. Introns must first be removed if eukaryotic DNA is to be expressed in a prokaryotic cell. The type of vector depends largely on the ultimate purpose of the procedure and the type of organism being engineered.
■ Explain the role of reverse transcriptase in the procedure to clone eukaryotic DNA.
■ Name three properties of an ideal vector.
■ Explain how the lacZ' gene is used to distinguish colonies that harbor a recombinant plasmid.
■ How could insertion of a vector randomly into the chromosome disrupt the regulation of existing genes?
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