Cloning and Expression of Thermus Aquaticus DNA Polymerase Gene, Using a Thermo-Inducible Expression Vector

DNA polymerase gene from Thermus aquaticus strain YT1 was amplified using VENTTM DNA polymerase and cloned under the control of X.PR promoter and expression was induced by a shift in tern perature. The culture was then sonicated, and after centrifugation the lysate was treated with polyethyleneimine followed by a salting-out step. Finally the protein was precipitated with ammonium sulfate and fractionated by gel filtration. The resulting enzyme preparation was stable and active.