麻风杆菌α2抗原基因的克隆及表达研究  

Gene Cloning and Expression of Mycobacterium Leprae α2 Antigen

作　　者：尹跃平[1] 铃木定彦[1] 牧野正直 吴勤学[1] 

机构地区：[1]中国医学科学院中国协和医科大学皮肤病研究所实验诊断研究室

出　　处：《中国医学科学院学报》1999年第1期62-68,共7页Acta Academiae Medicinae Sinicae

摘　　要：目的用现代分子生物学技术即基因工程手段制备麻风杆菌抗原。方法用原位斑点杂交法筛选阳性克隆，并对其中一株约３０００ｂｐ的ＤＮＡ片段进行酶谱和ＤＮＡ序列分析。结果从本实验组制备的麻风杆菌基因文库中筛选出２株含α２抗原的基因片段，并建立了α２抗原基因在大肠杆菌中的表达体系，在ＸＬＩ－Ｂｌｕｅ菌株中大量表达出易于提纯的α２重组融合蛋白抗原，经麦芽糖亲和色谱柱提纯后，每２００ｍｌ转化宿主菌培养液可获得１０ｍｇ重组融合蛋白。结论α２重组融合蛋白抗原的制备对于探讨麻风病的发病机制、建立麻风病和其它抗酸杆菌疾病（如结核病等）的高度特异性血清学诊断方法都有意义。Objectives The recombinant α2 antigen of M. leprae was prepared using the molecular biologic tools and the recombinant DNA expression technology.Methods Screening of the M leprae expression library was performed by the plaque hybridization technique. Nucleotide sequences were determined by dideoxy termination method. Results The gene coding for α2 antigen of M. leprae was cloned and characterized, and the complete nucleotide sequence data has been assigned in the GSDB, DDBJ, EMBL and NCBI nucleotide sequence databank. The over expression system of α2 antigen gene in E. coli was constructed, and the recombinant α2 antigen has been purified by amylose column chromatography at the purity of more than 95%. More than 10 mg of recombinant α2 antigen has been obtained from 200 ml of liquid culture. Conclusion The recombinant α2 antigen of M. leprae could be used as one of the specific antigens for the sero diagnosis of leprosy.

关 键 词：麻风杆菌 α2抗原 基因克隆 重组抗原 

分 类 号：R378.912[医药卫生—病原生物学] R755.03[医药卫生—基础医学]