Isolation and cloning of Rv3204 of Mycobacterium tuberculosis to Escherichia coli BL21 as vaccines tuberculosis : A preliminary Study

Abstract

Abstract Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis ( Mtb ). TB prevention is done through vaccination using the Bacille Calmette-Guerin (BCG) vaccine. There are weaknesses in this vaccine, so research is still needed in the search for specific antigens. One potential is the Rv3204 gene that encodes the methylated DNA-protein cycteine methyltransferase protein 11 kDa (MTSP11). This protein is immunogenic because it can produce interferon-γ (IFN-γ). The purpose of this study was to isolate and clone the Rv3204 Mycobacterium tuberculosis into Escherichia coli BL21 as vaccine tuberculosis. The research stage was amplification of Rv3204, ligation to pQE-30 Xa expression vector and transformation to Escherichia coli BL21 host cells on LB medium. Characterization was carried out by PCR analysis to recombinant plasmid isolation. The results showed that Rv3204 encoding MTSP11 was successfully cloned to E. coli BL21 with white colony formation and characterization showed that the band was 306 bp as the Rv3204 gene.

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