Table of Contents
Molecular Biology International
Volume 2014 (2014), Article ID 937308, 7 pages
http://dx.doi.org/10.1155/2014/937308
Research Article

Primer Based Approach for PCR Amplification of High GC Content Gene: Mycobacterium Gene as a Model

Department of Biotechnology, Panjab University, Sector 14, Chandigarh 160014, India

Received 21 August 2013; Revised 27 November 2013; Accepted 28 January 2014; Published 24 March 2014

Academic Editor: Alessandro Desideri

Copyright © 2014 Arbind Kumar and Jagdeep Kaur. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The genome of Mycobacterium is rich in GC content and poses problem in amplification of some genes, especially those rich in the GC content in terminal regions, by standard/routine PCR procedures. Attempts have been made to amplify three GC rich genes of Mycobacterium sp. (Rv0519c and Rv0774c from M. tuberculosis and ML0314c from M. leprae). Out of these three genes, Rv0774c gene was amplified with normal primers under standard PCR conditions, while no amplification was observed in case of Rv0519c and ML0314c genes. In the present investigation a modified primer based approach was successfully used for amplification of GC rich sequence of Rv0519c through codon optimization without changing the native amino acid sequence. The strategy was successfully confirmed by redesigning the standard primers with similar modifications followed by amplification of ML0314c gene.