Abstract

ISOLATION OF Taq POLYMERASE GENE & CLONING INTO E.coli USING pGEMT VECTOR

Meenakshi Lal and Sanjeev Ranjan*

Abstract

Use of the thermo stable Taq polymerase eliminated the need for having to add new enzyme to the Polymerase Chain Reaction (PCR) reaction during the thermo cycling process. A single closed tube in a relatively simple machine can be used to carry out the entire process. Thus, the use of Taq polymerase was the key idea that made PCR applicable to a large variety of molecular biology problems concerning DNA analysis.[1] Considering its huge role in molecular biology research the world market for Taq polymerase is in the hundreds of millions of dollars each year.[2] Gene cloning is the act of making copies of a single gene by introducing it in a host using a suitable vector. Amplified genes are useful in many areas of research and for medical applications such as gene therapy, isolation of genes for further manipulations, DNA fingerprinting, Molecular diagnostics, for cloning purpose leading to cheaper production of gene products, etc. Looking into the importance and ever increasing demands of Taq polymerase, it is important to focus on searching simpler methods for cloning it into easy to maintain host like Escherichia coli (E.coli). In the present study, cloning of Taq DNA Polymerase gene in E.coli host using pGEMT vector was done successfully.

Keywords: Taq DNA Polymerase, Polymerase Chain Reaction (PCR), Cloning, pGEMT vector