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  Recombinant DNA and Molecular cloning

Recombinant DNA:

The foreign DNA fragment isolated is made to recombine with plasmid DNA that is cleaved by similar restriction endonuclease. The recombination of the two DNAs is affected by the DNA ligase enzyme. The product created is termed as recombinant plasmid or recombinant DNA.

Molecular cloning:-

The recombinant DNA should be introduced into a host cell, inside which it might replicate freely. Escherechia coli have been used as an appropriate host to the above. Along with the multiplication and growth of the bacterium in the medium, copies of rDNA are also formed. In molecular cloning, alongside E.coli other microbes which have been used comprise Strptomyces sp., Bacillus subtilis, Saccharomyces cerevisieae and so on.

Whenever rDNA copies are introduced into host cells, (E. coli) a few thousand of rDNA pieces might enter the cells. Such cells are termed as transformed cells. Each transformed cell develops a colony of its own in which each and every member is genetically similar. Such colonies are then differentiated and recultured individually.

From the recultured colonies, the recombinant DNA is removed from lysed cells, purified and utilized. The initial gene was cloned in the year1973 by Hebert Bayer and Stanely cohen of Stanford University, California of USA.

 

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