Cloning and characterization of Rv1980c gene encoding MPT64 Protein from Mycobacterium tuberculosis as a new candidate vaccine of tuberculosis

Abstract

Abstract Pathogenic mycobacteria are one of the major causes of human mortality in the word. Mycobacterium tuberculosis is an etiological agent of human tuberculosis. Designing new vaccines including recombinant protein vaccines may be considered as a new approach for preventing or reducing tuberculosis epidemics. In order to construct protein recombinant as candidate vaccine, the Rv1980c gene encoding MPT64 protein was amplified from M. tuberculosis H37Rv strain genomic DNA using the PCR method and inserted into the cloning vector pGEM-T Easy. The recombinant plasmid pGEM-T Easy-MPT64 was then transformed into E. coli JM109 and cultivated under standard procedure, followed by plasmid extraction, PCR amplification, and DNA sequencing. The correct Rv1980c gene was confirmed by DNA sequencing and subcloned into expression vector pQE30Xa to yield recombinant plasmid pQE30Xa-MPT64, and transformed into E. coli BL21 strain. Transformed white recombinant colony was selected, cultured, induced with 40 μM IPTG, and identified using SDS-PAGE electrophoresis method. The molecular weight was found to be about 24 kDa and identified as recombinant protein MPT64. The target gene has been cloned into host E. coli BL-21 strain and expressed successfully as a soluble protein. The recombinant fusion recombinant protein MPT64 paves the way for tuberculosis diagnosis and vaccine development in the future, especially in Indonesia.