Cloning and expression of thermus aquaticus DNA polymerase gene, using a thermo - inducible experession vector

ABSTRACT

DNA polymerase gene from Thermus aquaticus strain YT1 was amplified using VENTTH DNA polymerase and cloned under the control of lambda pr promoter and expression was induced by a shift in temperature. 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