Cloning Open Reading Frame (ORF) of Rv2430c Mycobacterium tuberculosis Indonesian Isolate in Escherichia coli JM 109

2016 
Various strategies have been implemented to prevent tuberculosis. Vaccination with Bacille Calmette Guerin (BCG) vaccine is still used around the world. Generally, most people in have gained BCG vaccine as an infant, but the effectiveness of these vaccines do not survive to adulthood. Therefore, the necessary replacement BCG vaccine more effective to eliminate tuberculosis. Mycobacterium tuberculosis is an intracellular pathogen and it was inside the macrophage, which is considered to be the most important component of the immune system. M. tuberculosis has two sets of genes are highly polymorphic referred to as PE and PPE families. These unique families of proteins account for about 10% of the mycobacterial genome and have attracted great interest from a variety of different studies around the world. One member of the as a vaccine candidate is Rv 2430c. It is known that the sera of all patients infected with TB showed strong antibody responses against Rv 2430c compared to healthy individuals. The existence of these antibodies indicates that this protein is found in vivo during infection and is a native immunogenic molecule. The purpose of this study was to clone the Open Reading Frame (ORF) Rv 2430c M. tuberculosis Indonesian isolates to host cells E sherichia coli JM 109. The method used is by isolating -chromosomal DNA from clinical isolates from Indonesia, amplifying the ORF Rv 2430c with , ligating into cloning vectors pGEM-T and transform to E.coli JM 109. Characterization of clones do with migration analysis, restriction analysis and . The results obtained are recombinant clones that carry insertion was Rv 2430c Keywords: Mycobacterium tuberculosis, Rv 2430c, ORF, , JM 109 .
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