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All Journal Jurnal Penelitian Pendidikan IPA (JPPIPA)
Unsunnhidal, Lalu
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Education Materials Science & Nanotechnology Physics

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Cloning, expression and purification of a gene encoding CFP-10 and ESAT-6 protein of Mycobacterium tuberculosis in pET SUMO plasmids as Vaccine Candidates Fihiruddin, Fihiruddin; Inayati, Nurul; Srigede, Lalu; Zaetun, Siti; Unsunnhidal, Lalu; Shaleh, Muhammad Nazil
Jurnal Penelitian Pendidikan IPA Vol 12 No 2 (2026): In Progress
Publisher : Postgraduate, University of Mataram

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29303/jppipa.v12i2.12724

Abstract

The Bacillus Calmette-Guerin vaccine did not show consistent results in preventing tuberculosis, with an effectiveness ranging from 0% to 80%. More effective proteins are needed as vaccine candidates to eliminate tuberculosis. Culture filtrate proteins 10-kDa(CFP-10) and Excretory Antigen Target 6-kDa(ESAT-6) demonstrated very strong antigenicity against T and B cells. These proteins are secreted during the early phase of infection and could potentially be used for vaccine candidates. This study aims to clone and express the CFP-10 and ESAT-6 proteins on the pET SUMO Plasmid. PCR with specific primers was used to amplify the genes encoding CFP-10 and ESAT-6 proteins. The PCR products were cloned into pET SUMO plasmids for transformation into competent cells E.coli BL21(DE3). Recombinant protein expression was induced in LB medium using 1 mM IPTG. The yield of recombinant proteins was visualized by SDS-PAGE and western blotting using an Anti-HisTaq Antibody. PCR results showed the target gene sizes to be 304 bp (CFP-10) and 291 bp (ESAT-6). SDS-PAGE and Western blotting revealed proteins at 22kDa (CFP-10) and 18kDa (ESAT-6). The genes encoding the CFP-10 and ESAT-6 proteins of M. tuberculosis have been successfully cloned into the pET SUMO plasmid and expressed in E. coli BL21(DE3) with molecular weights corresponding to the target genes.
Co-Authors Fihiruddin, Fihiruddin Lalu Srigede, Lalu Nurul Inayati Shaleh, Muhammad Nazil Siti Zaetun, Siti