Prokaryotic Expression and Purification of Mouse Prilipin Gene
Objective To structure the prokaryotic expression vector of mouse prilipin gene, was expressed in prokaryotic cells and purify the expressed product. Methods By enzyme digestion from pMD18-Plin recombinant enzyme cut the prilipin gene.The target gene was recovered and cloned into expression vector pET-32a(+), and the constructed recombinant plasmid pET-32a-Plin was transformed to E.coli BL21(E3) for expression under induction of IPTG. The expressed product was identified by SDS-PAGE and Western blot, then purified by His label chromatography. Results The prokaryotic expression vector of mouse prilipin gene can express prilipin. The expressed recombinant prilipin, with a relative molecular mass of about 57 kDa, contained about 50% of total somatic protein. It showed good reactogenicity and reached a purity of more than 95% after purification. Conclusion The prokaryotic expression vector of mouse prilipin gene was successfully structured and expressed in prokaryotic cells, and the expressed product reached a high purity, which laid a foundation of study on the function of prilipin.
NETL
NSTL
万方数据
维普
