Open AccessCloning, expressing and purification of recombinant C-terminal PMT of Pasteurella multocida
Author(s) -
Hung K. Vu
Publication year - 2019
Publication title -
the journal of agriculture and development/the journal of agriculture and develoment/nông nghiệp và phát triển
Language(s) - English
Resource type - Journals
eISSN - 2615-949X
pISSN - 2615-9503
DOI - 10.52997/jad.8.05.2019
Subject(s) - pasteurella multocida , recombinant dna , pasteurellosis , microbiology and biotechnology , biology , escherichia coli , toxin , fusion protein , chemistry , bacteria , biochemistry , gene , genetics
Pasteurella multocida strains produce heat-stable toxin Pasteurella Multocida Toxin (PMT) which is the main virulence factor causing pasteurellosis in pigs. Therefore, PMT protein is considered as a promising candidate to study recombinant vaccine against pasteurellosis in pigs. In this study, we cloned the nucleotide sequence coding C-terminal fragment of PMT (tPMT-780), from acid amine 506 to acid amine 1285, into pRSET-A vector and expressed it on Escherichia coli BL21 bacteria. The SDS-PAGE result showed that tPMT-C780 was well-expressed by IPTG 1mM induction and determined to be insoluble protein, therefore it was purified by denaturation method using Ni-NTA beads. The protein elution samples contained only one protein band similar with the size of tPMT-C780 and positively reacted with PMT-specific antibody. These results suggested that we were successful in expressing and purifying C-terminal protein of PMT toxin, tPMT-C780. This is the primary material for further studies to produce recombinant vaccine against pasteurellosis in pigs.