in the synthesis of TaqI endonuclease using Ecoli strain by NEB company for example, where from they get the gene coded for this enzyme, what is the function of this enzyme in the original organism and by what technique can be extracted from E- coli?
An optimal purification procedure is dependent onquaticus, in the host organism, the enzyme provides defense from viruses and other invading DNA.
I do not know what methods NEB uses but Isolation of functional thermotolerant proteins from mesophilic host such as E.coli is quite simple and involves induction of foreign gene, followed by cell lysis, and heat inactivation and precipitation of host proteins. Since TaqI is a thermotolerant protein from a thermophile, it remains active after heat treatment.
Other proteins can be purified by incorporating His-Tag or S-Tag, if protein tagging hinders activity, the tag can be removed by thrombin cleavage or similar methods. An optimal purification procedure is dependent on the scale of production and the intended use.
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