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Enzymes & Vectors · Q9

Q.What is a cloning vector? Name the three essential features a plasmid must possess to function as a good cloning vector, and briefly explain the role of each.

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A cloning vector is a DNA molecule -- typically a bacterial plasmid or bacteriophage genome -- engineered to carry a foreign gene into a host cell and to be copied (replicated) inside that host along with the gene it carries. For a vector to work reliably as a cloning tool, it needs three essential features.

First, an origin of replication (ori): a specific DNA sequence at which the host cell's replication machinery begins copying the plasmid. Without an ori, the vector (and the foreign gene riding on it) would not be replicated when the host cell divides, and would simply be diluted out and lost. Second, a selectable marker gene, most commonly a gene conferring resistance to a particular antibiotic. This allows a researcher to grow treated host cells on a medium containing that antibiotic, killing off the vast majority of cells that never took up any vector DNA at all, and leaving only cells that did take up the vector. Third, one or more unique restriction sites, positioned …

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