中华皮肤科杂志 ›› 1996, Vol. 29 ›› Issue (6): 424-426.

• 论著 • 上一篇    下一篇

麻风杆菌MPL51蛋白基因的克隆及序列分析和表达

尹跃平1, 牧野正直2, 吴勤学1, 铃木定彦2   

  1. 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:1995-11-26 修回日期:1996-03-04 出版日期:1996-12-15 发布日期:1996-12-15

Cloning and Sequence Determination of MPL51 Protein of Mycobacterium Leprae

Yin Yueping1, Masanao Makino2, Wu Qinxue1   

  1. 1. Institute of Dermatology, Chinese Academy of Medical Sciences, Peking Union Medical Colege, Nanjing 210042;
    2. National Sanatorium Oku-Komyoen, Department, of Pathology, Osaka Prefectural Institute of Public Health
  • Received:1995-11-26 Revised:1996-03-04 Online:1996-12-15 Published:1996-12-15

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摘要： 麻风病的发病机理、疫苗的制备及早期血清学诊断的研究都需要提供高特异性的麻风杆菌抗原,而麻风杆菌体外尚不能人工培养,所以目前麻风杆菌的抗原分析和筛选主要依赖于分子生物学技术如基因工程等.MPT51蛋白是结核杆菌分泌蛋白的一种,具有多种免疫活性,且认为和85复合体抗原有同源性.本研究用原位斑点杂交法从麻风杆菌基因文库中筛选及克隆出编码麻风杆菌的MPT51类似蛋白即MPL51蛋白全基因片段,建立了此基因的酶切图谱,测定了全基因的DNA顺序.并在大肠杆菌中成功地表达出分子量为38000的重组蛋白,用蛋白印迹试验初步检测表明,在麻风患者血清中存在抗表达产物MPL51蛋白抗体.结果表明麻风杆菌MPL51蛋白可能同源于85复合体蛋白,有可能成为在麻风病的发病机理和血清学诊断方法等研究中的重要抗原之一.

关键词: 麻风分支杆菌, 基因文库, MPL51蛋白

Abstract: The MPT51 protein is one of the secreted proteins from Mycobacterium tuberculosis and its N-terminal amino acid is reported to have about 60% homology with 85B antigen.In this study,we have characterized the MPL51 protein from Mycobacterium leprae which is similar to MPT51 protein.We have sequenced 3kbp DNA fragment which involves MPL51 opening fragment and 85 A antigen gene fragment.In the study of analysis of N-terminal amino acid sequence,it has 80% homology with MPT51 and 57% with 85A,Band C respectively.By the results of Western blotting,we found that antibodies agains tMPL51 protein existed in the sera of leprosy patients and also in healthy control sera no other antibody was found.

Key words: Mycobacterium leprae, Genomiclibrary, MPL51 protein