Abstract:
Objective To construct the prokaryotic expression vector of NFA47650 protein from Nocardia farcinica, induce the expression and purification of NFA47650 protein, and analyze its immunogenicity.
Methods The primers were designed and synthesized based on nfa-47650 gene sequence of N. farcinica; pET30a (+) expression vector was constructed by PCR amplification; NFA47650 protein was purified after expressing in E. coli BL21. The subcellular localization and immunogenicity with NFA47650 mice anti-serum were analyzed by Western Blot.
Results The prokaryotic recombinant expression vector of N. farcinica NFA47650 protein was successfully constructed, which was efficiently expressed in BL21 E. coli as inclusion bodies. Western Blot showed that NFA47650 protein was mainly located in the culture supernatant. In addition, this protein could not only be specifically identified by the antisera from mice and rabbits infected with N. farcinica, but also cross-reacted with antisera from mice infected with N. brasiliensis and N. cyriacigeorgica.
Conclusion The NFA47650 protein is an important secreted protein, which can trigger immune response by reacting with the antisera of different Nocardia species.
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