Isolation And Cloning Of Gag-Ca Gene Of The Jembrana Disease Virus On Pgex-2T Procaryotic Expression Vector

Jembrana disease virus (JDV) is agent of an acute infectious disease of Bali cattle. Gag protein is one of the most immunogenic structural proteins. This work wan intended to construct a procaryotic expression vector based on pGEX-2T for recombinant protein production in E. coli. Gag-Ca gene was excised from our previous construct pCR-C A with BamHI-EcoRI and inserted in these sites in pGEX-2T. With good primer concept for the gene isolation and the procedures used for cloning, cloning efficiency in pGEX-2T and transformation in E. coli DH 5 were very high. There are about 1,500-2,000 recombinant bacterial colonies were obtained. Cloning efficiency in pGEX-2T attained 100%. Finally, the conception of the construction also resulted in sens oriented-insertion so that correct protein synthesis will take place. Keywords: Jembrana virus, pGEX-2T, Recombinant protein, Gag-Ca gene