绵羊肺炎支原体新疆分离株黏附素与溶血素A基因的克隆及序列分析  

Cloning and Molecular Characteristics of Adhesin and Hemolysin A Gene of Mycoplasmao vipneumoniae

作　　者：胡政香 乔军[1] 孟庆玲[1] 陈诚[1] 赵海龙[1] 马玉[1] 刘田莉 赞哈尔.波拉提 张金生 陈创夫[1] 

机构地区：[1]石河子大学动物科技学院,石河子832003 [2]新疆沙湾县兽医站,沙湾834700

出　　处：《石河子大学学报（自然科学版）》2015年第2期201-205,共5页Journal of Shihezi University(Natural Science)

基　　金：国家国际科技合作专项(2014DFR31310);新疆兵团国际科技合作计划项目(2012BC006)

摘　　要：根据Gen Bank报道的绵羊肺炎支原体(MO)基因组序列,设计特异性引物,对MO新疆分离株黏附素基因和溶血素A基因进行PCR扩增、克隆及测序;应用分子生物学软件对该基因及其编码蛋白进行分子特征分析,并与其他支原体相应序列进行同源性分析,构建该基因系统进化树。结果显示:黏附素基因长为3426 bp,溶血素A基因长为777 bp;黏附素在第1-23位氨基酸残基为信号肽、1个跨膜区,可能为分泌型蛋白;溶血素A无信号肽和跨膜区、有RNA结合位点和甲基转移酶结合位点。系统发生分析表明,黏附素基因与猪肺炎支原体P146株进化关系较近,与其他的支原体进化关系远;溶血素A基因与猪肺炎支原体232株、结膜支原体HRC581株位于同一分枝上,进化关系近。本研究克隆了MO黏附素基因和溶血素A基因,为了解MO毒力相关基因生物学功能及其致病中的作用奠定了前期基础。According to the genomic sequence of Mycoplasmao vipneumoniae （MO）,the adhesin and hemolysin A gene of MO Xinjiang isolate（XJ-MO）were amplified with designed specific primers by PCR,and then sequenced.The molecular characteristic of Adhesin and hemolysin A gene and their encoding protein were analyzed by molecular biology software,and the sequences of Adhesin had a high among other isolates of Mycoplasmao vipneumoniae was analyzed.Phylogenetic tree was then constructed to analyze the relationship among different Mycoplasma.The results showed that the Adhesin gene and hemolysin A gene of XJ-MO strain contains 3426 bp and 777 bp respectively.The Adhesin deduced protein had a signal peptide sequence between amino acids 1 and 23,a transmembrane region,may be secreted proteins;hemolysin A no signal peptide and transmembrane region,which had RNA binding sites and Methyltransferase binding site.Polypeptide chain is mainly composed of random curl.It is inferred that adhesin is hydrophobic protein secretion.Phylogenetic tree identities with Mycoplasma hyopneumoniae P146 and the far distant with other Mycoplasma strains;hemolysin A with Mycoplasma hyopneumoniae 232 had a high identities MO adhesin and hemolysin A genes was first cloned in this study,which laid a foundation for the understanding of biological functions of Virulence gene and it＇s role in pathogenicity.

关 键 词：绵羊肺炎支原体 黏附素 溶血素A 基因克隆 序列分析 

分 类 号：S852.62[农业科学—基础兽医学] S811.6[农业科学—兽医学]