Jurnal Penelitian Pendidikan IPA (JPPIPA)
Vol 12 No 2 (2026): In Progress

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 (Unknown)
Inayati, Nurul (Unknown)
Srigede, Lalu (Unknown)
Zaetun, Siti (Unknown)
Unsunnhidal, Lalu (Unknown)
Shaleh, Muhammad Nazil (Unknown)



Article Info

Publish Date
28 Feb 2026

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.

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Journal Info

Abbrev

jppipa

Publisher

Subject

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Education Materials Science & Nanotechnology Physics

Description

Science Educational Research Journal is international open access, published by Science Master Program of Science Education Graduate Program University of Mataram, contains scientific articles both in the form of research results and literature review that includes science, technology and teaching ...