鸭疫里默氏菌贵州分离株OmpA基因的克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of OmpA Gene of Riemerella anatipestifer Guizhou Isolate

作　　者：马光强 刘丽娟 陈国权 吴良涛 杨均[1] 刘军泽 李潇蒙 潘成文 周碧君[3] MA Guang-qiang;LIU Li-juan;CHEN Guo-quan;WU Liang-tao;YANG Jun;LIU Jun-ze;LI Xiao-meng;PAN Cheng-wen;ZHOU Bi-jun(Qiannan Polytechnic College for Nationality,Duyun,Guizhou,558000,China;Agriculture and Rural Affairs Bureau of Duyun,Duyun,Guizhou,558000,China;College of Animal Science,Guizhou Uiversity,Guiyang,Guizhou,550025,China)

机构地区：[1]黔南民族职业技术学院,贵州都匀558000 [2]都匀市农业农村局,贵州都匀558000 [3]贵州大学动物科学学院,贵州贵阳550025

出　　处：《动物医学进展》2022年第9期12-19,共8页Progress In Veterinary Medicine

基　　金：三穗鸭工程技术研究中心建设项目(黔科合平台人才[2019]5203号);贵州省科技平台及人才团队计划项目(黔科合平台人才[2018]5253号)。

摘　　要：为了解鸭疫里默氏菌(Riemerella anatipestifer,RA)贵州分离株OmpA基因遗传特性和用作基因工程亚单位疫苗开发的可能性,以13株RA贵州分离株(RA-SS-1~12株和RA-AS-1株)为研究对象,用RA OmpA基因特异性引物对13株RA贵州分离株进行PCR扩增,将扩增得到的OmpA基因片段与pMD19-T载体连接,转化大肠埃希氏菌DH5α感受态细胞,经PCR和双酶切鉴定为阳性克隆后进行测序;用生物信息学软件将13株RA贵州分离株与国内外15株参考株OmpA基因序列进行核苷酸同源性分析和遗传进化分析,并对RA OmpA蛋白进行理化性质分析、跨膜结构和信号肽预测、B细胞抗原表位预测、二级和三级结构预测。结果表明,13株RA贵州分离株OmpA基因全长均为1164 bp,可编码387个氨基酸,株间核苷酸同源性为100%,与参考株GZ-RA8、GZ-RA9、GZ060302、SD-RA2018和Yb2株的同源性达100%”;RA OmpA蛋白属亲水性蛋白,无跨膜和信号肽区域,含有较多的优势抗原表位结构,且OmpA基因高度保守并具有良好的抗原特性,其可作为制备RA基因工程亚单位疫苗的候选蛋白。This article aims to understand the genetic characteristics of the OmpA gene of Riemerella anatipestifer(RA)Guizhou isolate and the possibility of using it as a genetic engineering subunit vaccine.13 RA Guizhou isolates(RA-SS-1-12 strains and RA-AS-1 strains)were the research objects.13 RA Guizhou isolates were amplified by PCR with RA OmpA gene-specific primers.The amplified OmpA gene fragment was connected to the pMD19-T vector,transformed it into E.coli DH5a competent cells.The positive clones was identified by PCR and double enzyme digestion,and then sequenced;The OmpA gene sequences of 13 RA Guizhou isolates and 15 domestic and foreign reference strains were analyzed for nucleotide homology using bioinformatics software and genetic evolution analysis,the physical and chemical properties analysis,transmembrane structure and signal peptide prediction,B cell epitope prediction,secondary and tertiary structure prediction of RA OmpA protein were conducted.The results showed that the full length of OmpA gene of 13 RA Guizhou isolates was 1164 bp,which could encode 387 amino acids.The nucleotide homology between the strains was 100%,and the nucleotide homology to the reference strain RA GZ-RA8、GZ-RA9、GZ060302、SD-RA2018 and Yb2 was 100%.RA OmpA protein is a hydrophilic protein with no transmembrane and signal peptide regions,and contains more dominant epitope structures,and the OmpA gene is highly conservative and has good antigenic characteristics.It is predicted that it can be used as a candidate protein for genetic engineering sub-unit RA vaccine preparation.

关 键 词：鸭疫里默氏菌 OmpA基因 生物信息学分析 

分 类 号：S852.6[农业科学—基础兽医学]