麻风杆菌α-抗原C基因的克隆及表达

中华皮肤科杂志 ›› 1996, Vol. 29 ›› Issue (5): 343-345.

麻风杆菌α-抗原C基因的克隆及表达

尹跃平¹, 牧野正直², 吴勤学¹, 铃木定彦²

1. 中国医学科学院、中国协和医科大学皮肤病研究所, 南京210042;
2. 日本大阪大学公众卫生研究所

收稿日期:1995-11-21 修回日期:1996-03-04 出版日期:1996-10-15 发布日期:1996-10-15
基金资助:
日本Sasakawa纪念保健协力财团麻风研究基金资助课题

Construction of Recombinant 85C Antigen of Mycobacterium Leprae in E.coli

Yin Yueping¹, Masanao Makino², Wu Qinxue¹

Institute of Dermatology, Chinese Academy of Medical Sciences, Peking Union Medical College, Nanjing 210042

Received:1995-11-21 Revised:1996-03-04 Online:1996-10-15 Published:1996-10-15

摘要/Abstract

摘要： 在分支杆菌主要分泌蛋白之一的α-抗原组中,目前已知的有三种蛋白,即85A、B和C抗原。我们已报道了麻风杆菌α-抗原85A和B基因的克隆以及在大肠杆菌中的表达。为了筛选出用于麻风病早期血清学诊断以及病因探讨的特异性高和免疫性强的抗原,我们又用聚合酶链反应技术克隆出麻风杆菌α-抗原C基因,并将此基因插入表达载体pT7HDXa、转化宿主菌XLI-Blue,以IPTG及M13TTpol作为诱导剂,诱导表达α-抗原C的重组蛋白,检测结果表明麻风杆菌α-抗原C抗原在此表达体系中能够进行表达,这将有助于进一步开展对α-抗原组开发应用的研究。

关键词: 麻风杆菌, 抗原85复合体C

Abstract: The α-antigen is one of the dominant mycobacterial protien and consists of three structurally related components which are designated as 85A,85B and 85C.We have reported the molecular cloning of 85A and 85B of M.leprae and its over production in E.coli.In order to screen an antigen with high specificity and immunnity for the serodiagnosis and explore the etiology of leprosy.We continue to study the construction of the 85C antigen of Mycobacterium leprae in E.coll.M.leprae 85C gene was amplified by PCR technique using primers 5′-terminus and 3′-terminus which contain BamH Ⅰ and Hind Ⅲ recognition sequence,respectively,and insert amplified fragment into expression vector. Ampicillin resistant transformats were cultured in LB medium and inducedby IPTG and M13T7pol to produce M.leprae 85C fusion protein.The recombinant 85C protein was recognized by anti-α-TB antisera.The result of this study will be useful for the further study of a-antigen.

Key words: Mycobacterium leprae, Antigen 85 complex

尹跃平, 牧野正直, 吴勤学, 铃木定彦. 麻风杆菌α-抗原C基因的克隆及表达[J]. 中华皮肤科杂志, 1996, 29(5): 343-345.

Yin Yueping, Masanao Makino, Wu Qinxue. Construction of Recombinant 85C Antigen of Mycobacterium Leprae in E.coli[J]. Chinese Journal of Dermatology, 1996, 29(5): 343-345.

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