中国畜牧兽医 ›› 2014, Vol. 41 ›› Issue (9): 59-62.

• 生物技术 • 上一篇    下一篇

牛支原体不同分离株全菌蛋白免疫印迹分析

陈忠琼1,2, 范媛1,3, 蒋勇1, 周蓓1, 黎晓敏1   

  1. 1. 西南大学动物科技学院, 重庆 400715;
    2. 重庆市动物疫病预防控制中心, 重庆 401120;
    3. 重庆市万州动物卫生监督所, 重庆 404000
  • 收稿日期:2014-01-13 出版日期:2014-09-20 发布日期:2014-09-24
  • 通讯作者: 黎晓敏 E-mail:lixiaomin662@sina.com
  • 作者简介:陈忠琼(1987- ),女,贵州人,硕士生,研究方向:疾病病理学。
  • 基金资助:

    重庆市科委重大攻关项目(CSTC,2010AA1021)。

Western Blotting Analysis on the Whole Proteins of Different Isolates of Mycoplasma bovis

CHEN Zhong-qiong1,2, FAN Yuan1,3, JIANG Yong1, ZHOU Bei1, LI Xiao-min1   

  1. 1. College of Animal Science and Technology, Southwest University, Chongqing 400715, China;
    2. Chongqing Center for Animal Disease Control and Prevention, Chongqing 401120, China;
    3. Animal Health Supervision Institute of Wanzhou, Chongqing 404000, China
  • Received:2014-01-13 Online:2014-09-20 Published:2014-09-24

摘要: 为比较牛支原体(Mycoplasma bovis,M.bovis)不同分离株间全菌蛋白组成的差异,找到其具有免疫原性的蛋白片段,试验采用裂解液法提取分离自全国不同地区6株M.bovis分离株(W70株、1738株、Q3株、Q1株、JX02株、677株)的全菌蛋白,并利用自制抗血清对所获蛋白进行SDS-PAGE和免疫印迹分析。结果显示,6株分离株全菌蛋白条带数量、清晰度存在差异,其中W70株、1738株、Q3株和Q1株的蛋白条带数量及清晰度均优于JX02株和677株,蛋白质分子质量范围在23.2~130.8 ku之间;6株分离株均显现2条大小为55和43 ku的免疫杂交条带。综上所述,M.bovis不同分离株全菌蛋白组成存在差异,55和43 ku是其主要的免疫原性蛋白之一,该试验结果为M.bovis病血清学诊断、分子诊断技术及疫苗的研制提供理论依据。

关键词: 牛支原体; 全菌蛋白; SDS-PAGE; 免疫印迹

Abstract: The whole proteins of 6 Mycoplasma bovis (M.bovis) isolates (W70 isolate,1738 isolate,Q3 isolate,Q1 isolate,JX02 isolate and 677 isolate),which isolated from different parts of the country was extracted by lysate,were analysed by SDS-PAGE and Western blotting with homemade antiserum.The results showed that there were differences on polypeptide quantity and definition of the whole proteins of 6 isolates:W70 isolate,1738 isolate,Q3 isolate and Q1 isolate were better than JX02 isolate and 677 isolate,which molecular weight were 23.2 to 130.8 ku.6 isolates were appeared 55 and 43 ku band when analysed by Western blotting.In conclusion,there were differences on polypeptide composition of whole proteins of the different M.bovis isolates,and 55 and 43 ku were the main immunogenicity protein of M.bovis.These results provided the theoretical basis for serology diagnosis,molecular diagnosis and the vaccine development of the M.bovis disease.

Key words: Mycoplasma bovis; whole proteins; SDS-PAGE; Western blotting

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