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Surong Hasi

Surong Hasi

Inner Mongolia Agricultural University, China

Title: Prokaryotic expression and purification of CYP2J protein of Bactrian camel


Biography: Surong Hasi


CYP2J enzyme plays a very important role in the metabolism of endogenous and exogenous compounds. The purpose of this experiment is to express CYP2J protein of the Bactrian camel and purify it. Firstly, based on the CYP2J gene sequence of Bactrian camel and the preference of E. coli codons, codon of CYP2J gene were optimized and was synthesized. Then, the recombinant expression vector pET-21a-CYP2J was constructed and transformed into E. coli BL21 (DE3) competent cells. After recombinant vector was induced by IPTG, the optimal expression conditions were determined by investigating induction temperature and induction time. Finally, the expressed product was purified by Ni-NTA Sepharose Affinity Chromatography and identified by SDS-PAGE and Western blot. The results showed that the recombinant expression vector of PET-21a-CYP2J was successfully constructed. And the best expression condition of CYP2J protein was IPTG concentration of 0.5 mmol/L, induction temperature of 20℃, and induction time of 8 h. The recombinant protein about 55 kD was detected by SDS-PAGE, and the Western blot analysis further proved that the CYP2J protein was expressed successfully. In conclusion, the successful expression of Bactrian camel CYP2J protein laid the foundation for the next monoclonal antibody preparation and determination of Bactrian camel CYP2J enzyme expression and distribution at the protein level.