Competent Host (For Transformation with Recombinant DNA)

The propagation of the recombinant DNA molecules must occur inside a living system or host. Many types of host cells are available for gene cloning which includes E.coli, yeast, animal or plant cells. The type of host cell depends upon the cloning experiment. E.coli is the most widely used organism as its genetic make-up has been extensively studied, it is easy to handle and grow, can accept a range of vectors and has also been studied for safety. One more important feature of E.coli to be preferred as a host cell is that under optimal growing conditions the cells divide every 20 minutes. Since the DNA is a hydrophilic molecule,it cannot pass through cell membranes, In order to force bacteria to take up the plasmid, the bacterial cells must first be made competent to take up DNA.

This is done by treating them with a specific concentration of a divalent cation such as calcium. Recombinant DNA can then be forced into such cells by incubating the cells with recombinant DNA on ice, followed by placing them briefly at 420C (heatshock) and then putting them back on ice.

This enables bacteria to take up the Recombinant DNA. For the expression of eukaryotic proteins, eukaryotic cells are preferred because to produce a functionally active protein it should fold properly and post translational modifications should also occur, which is not possible by prokaryotic cell (E.coli).

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