解脲脲原体MB蛋白在大肠埃希菌中的表达与纯化

Expression and Purification of the Multiple Banded Proteins of Ureaplasma Urealyticum in Escherichia coli

目的在大肠埃希菌中克隆、表达解脲脲原体种特异性抗原（MB）蛋白并纯化出重组蛋白.方法利用Gateway系统将MBA蛋白5’端保守性序列克隆到表达载体pDEST17上，并通过载体自身所带6XHis标签，利用Ni—NTA纯化出相应的重组蛋白.结果MB蛋白5’端414bp的保守序列成功地克隆到pDET17载体上并且表达出相应大小的目的蛋白.通过Ni—NAT纯化出重组表达的MB蛋白，其纯度达到93%，浓度为0．6mg/ml.结论解脲脲原体的MB蛋白的体外表达，为下一步制备相应抗体从而建立解脲脲原体的血清学检测方法打下基础.

Objective To express and purification the species-specific protein of Ureaplasma urealyticum, the multiplebanded antigen. Methods Cloning the 5＇end of the multiple banded antigen genes into expressing vector pDEST17 by Gateway system and purifying the recombinant protein by Ni-NAT. Results The 5＇end of the multiple-banded antigen gene was successfully cloned into the vector pDEST17 and the appropriated molecular weight protein was expressed in E. coli BL21（DE3）. The purity of the purified recombinant MBA protein was about 93% and the concentration was 0.6 mg/ ml. Conclusion The expression of MBA protein in E. coli gives the basic to prepare antibody against MBA and establish the serodiagnostic method for detecting the infection of Ureaplasma urealyticum expediently and rapidly.