Cloning in Cosmid Vectors, Plasmid, Pieces of DNa, Concatameric Molecules, DNA Ligase, Chimeric Cosmid DNA, Circularziers, Phage Genome

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Cloning in cosmid vectors

Cosmids were developed to allow cloning of large DNA fragments (-40 kb) in plasmid/phage λlike vectors. The vector is cut with a restriction enzyme and mixed with pieces of DNA to be cloned The cleaved vector and the foreign DNA fragments are made to join to produce concatameric molecules with the help of DNA ligase (concatamers = several molecules joined together end to end).

The ligation mixture is then mixed with a packaging extract (commercially available) to allow packaging of chimeric cosmid DNA into phage heads. The cosmid containing phages are infectious. After infection, the cosmid circularizes, and multiplies in E. colicells, just like a plasmid.

Like a plasmid, a cosmid is also small in size (5-8 kb), but like a phage it can be packaged in λ head, thus permitting cloning of large DNA fragments (-40 kb; λ phage genome = 50 kb and therefore can accommodate no more then -20 kb).

Cloning of a large DNA fragment in a cosmid vector

Cloning of a Large DNA Fragment in a Cosmid Vector

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