Journal of Pure and Applied MicrobiologyVol. 9 No. Special Edition May. 2015

Molecular Characterization of ORF esat Gene of M. tuberculosis Isolated from E.coli DE3

Chang-zhong Liu, Guang-hui Wei, Jian-he Hu and Xing-you Liu*

College of Animal Science, Henan Institute of Science and Technology, Xinxiang, Henan Province, China.

Received on 03 January 2015 and accepted on 06 February 2015

 

ABSTRACT

In this study, the open reading frame of a Secretory Antigenic Target (Esat) gene from M. tuberculosis in Escherichia coli BL21 (DE3) was cloned. Esat gene that was isolated from pulmonary TB patient was cloned into a plasmid vector (pGEM-Teasy) to construct pMB38. The E.coli DE3 clone carrying pMb38 was selected on X-gal medium. The expression of ORF in Esat gene was mediated using pRoExHTc under the control of Tre promoter and E.coli DE3 as host. Characterization of clones with recombinant plasmid was cut with restriction enzymes and sequencing of DNA inserts. Our results by BLAST analysis of sequencing showed 100% homology. It is hoped these proteins can then be used as an antigen for serological detection of latent TB in people.

Keywords : ESAT gene; Clone; M. tuberculosis; E.coli.