Cloning,expression and purification of Nocardia brasiliensis proteion P61 with biological activity
JI Xing-zhao, TANG Lu, HOU Xue-xin, SUN Li-na, WEI Chao, XU Shuai, SI Chen-chen, LI Zhen-jun
Institute for Communicable Disease Prevention and Control,Stat-Key Laboratory for Communicable Disease Prevention and Control,Chinese Center for Disease Control and prevention, Beijing 102206,China
Abstract:We constructed prokaryotic recombinant expression vector of P61 gene from Nocardia brasiliensis, expressing P61 protein with biological activity in E.coli, and lay a foundation for further studies related to P61. P61 gene was synthesized and cloned into an expression vector pET-30a(+).The recombinant vector was transformed into Escherichia coli BL21 and induced with IPTG. The production was analyzed with Western blot and the catalase activity of P61 was tested with Catalase Assay Kit. The protein of P61was successfully expressed in E. coli with solubility and high catalase activity, and could be identified by anti-N. brasiliensis sera from mice. The prokaryotic expression plasmid of protein P61 was constructed successfully and can be expressed efficiently in E.coli BL21 cells with higher catalase.
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