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Home » Biology Homework Help » Biotechnology » Coli Vectors and Plasmids
Coli Vectors and Plasmids
Coli vectors

Bacteria are the hosts of choice for DNA cloning. Among then, E. coli occupies a prominent position since cloning and isolating DNA inserts for structural analysis is the easiest in this host. Therefore, the initial cloning experiments are generally carried out in the E. coli. The E. coli strain K12 is the most commonly used; it has several substrains, e.g. C 600, RRI, HB 101 etc. each of which has some specific features important in cloning. For example, the substrain RRI has, in addition to certain other features, the mutation hsdR which inactivates the restriction enzyme endogeneous to E. coli K 12; this degradation of recombinant DNA introduced into it.

Properties of a good host

A good host should have the following features: (1) is easy to transform, (2) supports the replication of recombinant DNA, (3) is free from elements that interfere with replication of recombinant DNA, (4) lacks active restriction enzymes, e.g. E. coli K12 substrain HB 101, (5) does not have methylases since these enzymes would methylate the replicated recombinant DNA which, as a result, would become resistant to useful restriction enzymes, and (6) is deficient in normal recombination function so that DNA insert is not altered by recombination events.

Plasmids

A plasmid is a DNA molecule, other than the bacterial chromosome, that is capable of independent replication and transmission. Plasmids are circular and may exist either independent of or may become integrated into the bacterial chromosome; generally they are not essential for the host cell except under specific environments. There are several types of bacterial plasmids, but the three widely studied types are: F plasmids (responsible for conjugation), R plasmids (carry genes for resistance to antibiotics) and Col plasmids (code for colicins, the proteins that kill sensitive E. coli cells; they also carry genes that provide immunity to the particular colicin). The plasmids may either be conjugative or transmissible (mediate DNA transfer through conjugation, and as a result spread and some of Col plasmids, or non conjugative (do not mediate DNA transfer through conjugation) e.g. many R plasmids and most Col plasmids.

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