广东与四川地区发病猪副猪嗜血杆菌分离株omp5基因的克隆及序列分析  

Clone and sequence anlysis of outer membrane protein P5 gene of Haemophilus parasuis isolated from healthy and disease swine

作　　者：陈善真[1,2,3] 李春玲[2,3] 王贵平[2,3] 曾振灵[1] 贾爱卿[2,3] 杨冬霞[2,3] 

机构地区：[1]华南农业大学兽医学院,广东广州510642 [2]广东省农业科学院兽医研究所,广东广州510640 [3]广东省兽医公共卫生实验室,广东广州510640

出　　处：《黑龙江畜牧兽医》2011年第3期12-16,共5页Heilongjiang Animal Science And veterinary Medicine

基　　金：广东省农业科技重点引导项目(2007A020300005-11);广东省农业科技重大专项(2009A020101006);广州市农业科技重大专项(2009A1-E041)

摘　　要：为了比较副猪嗜血杆菌(HPS)外膜蛋白P5(omp5)基因的同源性及抗原性,试验设计合成1对特异性引物,通过PCR方法从广东省和四川省发病猪的30株分离菌中扩增出HPS omp5基因,然后将PCR产物克隆至pMDTM18-T载体上,转化大肠杆菌DH5α,并进行序列测定与分析。结果表明:30株HPS omp5基因的完整序列有3种长度,分别为1 116 bp、1 104 bp、1 098 bp;与已发表的血清5型HPS SH0165的核苷酸序列及其推导的氨基酸序列比较,广东分离菌株核苷酸同源性为90.5%～100%,氨基酸同源性为88.8%～100%;而四川分离菌株的核苷酸序列同源性为90.8%～94.1%,氨基酸同源性为89.9%c93.0%;进化树分析显示,HPS omp5基因无论是在不同血清型之间还是不同地区之间都具有较高的同源性,说明OMP5蛋白比较保守,有望作为型间共用性抗原。To compare the homology and antigenicity of outer membrane protein P5（omp5） gene between different serotypes of Haemophilus parasuis（HPS）,a pair of primers was designed according to Haemophilus parasuis outer membrane protein gene sequences published in the GenBank.The omp5 genes of 30 strains were amplified by PCR,in which 16 strains belonging to three serotypes were isolated from ill pigs in Guangdong and the other 14 strains were from Sichuan.The amplified fragments were inserted into the pMD18-T vector respectively,then sequenced and analyzed.The results showed that the omp5 genes were 1 116 bp,1 104 bp and 1 098 bp in size,encoding 372,368 and 366 amino acid residues respectively.The homology of nucleotide sequence of Guangdong strains with published Haemophilus parasuis SH0165 outer membrane protein gene were 90.5%～100%,and 88.8%～100% at amino acid level.The homology of nucleotide sequence from Sichuan strains with SH0165 were between 90.8%～94.1%,and 89.9%～93.0% at amino acid level.Phylogenetic analysis also showed that the,omp5 genes of HPS had high homology among the different serotypes,or between different regions.It indicates that the OMP5 protein of HPS are more conservative and used as a general antigen.

关 键 词：副猪嗜血杆菌 omp5基因 克隆 序列比较 

分 类 号：S852.612[农业科学—基础兽医学]