我国17株绵羊肺炎支原体分子分型及其菌体蛋白免疫印迹  

Molecular typing and immunoblotting of 17 Mycoplasma ovipneumoniae isolates from China

作　　者：许健[1,2] 储岳峰[1] 高鹏程[1] 赵萍[1] 贺英[1] 剡根强[2] 逯忠新[1] 

机构地区：[1]中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,农业部草食动物疫病重点开放实验室,农业部兽医公共卫生重点开放实验室,甘肃省生物检测工程技术研究中心,兰州730046 [2]石河子大学动物科技学院,石河子832000

出　　处：《微生物学报》2011年第10期1421-1426,共6页Acta Microbiologica Sinica

基　　金：甘肃省科技支撑计划项目(1011NKCA054);甘肃省农业生物技术专项(GNSW-2010-09);国家科技基础性工作专项(2008FY210200);国家现代肉羊产业技术体系(CARS-39)~~

摘　　要：【目的】分别从基因和蛋白水平研究我国部分地区绵羊肺炎支原体(Mycoplasma ovipneumoniae)分离株的分子特征,并了解其免疫原性蛋白的差异。【方法】对分离自8个地区的17株绵羊肺炎支原体进行扩增片段长度多态性(Amplified Fragment Length Polymorphism,AFLP)和十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis,SDS-PAGE)分析,采用NTsys-2.10e软件对AFLP和SDS-PAGE结果进行聚类,并用绵羊肺炎支原体模式株Y98高免血清对部分分离株进行免疫印迹分析。【结果】当相似系数分别为0.78和0.85时,绵羊肺炎支原体分离株可根据8个来源地区分成8个AFLP群和8个SDS-PAGE群;用8株分离株进行免疫印迹共出现6条蛋白条带,分子质量分别为105 kDa、83 kDa、65 kDa、42 kDa、40 kDa或26 kDa,其中83 kDa和40 kDa蛋白为8个菌株保守的免疫原性蛋白。【结论】我国部分地区绵羊肺炎支原体分离株之间存在遗传差异,不同分离株的主要免疫原性蛋白也存在一定差异,但83 kDa和40 kDa蛋白为其保守的免疫原性蛋白。本研究首次对我国部分地区绵羊肺炎支原体分离株进行了分子分型与免疫印迹分析,结果将为绵羊肺炎支原体病的新型诊断技术开发和疫苗研制奠定理论基础。[Objective] To study the heterogeneity and immunogenic variability among Mycoplasma ovipneumoniae（M.ovipneumoniae） isolates from differnt regions of China.[Methods] The heterogeneity of 17 strains of M.ovipneumoniae isolated from 8 regions of China was studied by the amplified fragment length polymorphism（AFLP） and sodium dodecyl sulphate-polyacrylamide gel electrophoresis（SDS-PAGE）.The software NTsys-2.10e was used to analyze the profiles obtained from the AFLP and SDS-PAGE.The proteins reacted with the antiserum against M.ovipneumoniae type strain Y98 were then detected by Western-blot.[Results] Seventeen strains of M.ovipneumoniae were divided into 8 AFLP groups based on the source regions when the coefficient was 0.78.They were also divided into 8 SDS-PAGE groups based on the source regions when the coefficient was 0.85.A total of 6 immunogenic proteins were detected within 8 strains of M.ovipneumoniae,and their molecular weights were 105 kDa,83 kDa,65 kDa,42 kDa,40 kDa or 26 kDa,respectively.Interestingly,the 83 kDa and 40 kDa proteins were conserved in all the 8 isolates.[Conclusion] M.ovipneumoniae isolates from some regions of China were genetically different,but the 83 kDa and 40 kDa antigenic proteins were conserved among the tested isolates.This study can provide some insights for the diagnosis and vaccine development of the disease caused by M.ovipneumoniae.

关 键 词：绵羊肺炎支原体 AFLP SDS-PAGE 免疫印迹 

分 类 号：S858.26[农业科学—临床兽医学]