牛乳腺炎金黄葡萄球菌弹性纤维结合蛋白N端蛋白基因的克隆及序列分析  

Cloning and sequencing of the gene (nEBPS) for the N-terminal domain of the Streptococcus aureus fibronectin-binding protein from cows with mastitis

作　　者：薛晓阳[1,2,3] 吴金花[1,2] 布日额[1,2] 王学理[2,3] 刘洋[1,2,3] 刘燕[1,2] 锡林高娃[1,2] 孙立杰[1,2] 郭闯[1,2] 薛江东[2,3] 

机构地区：[1]内蒙古民族大学生命科学学院,内蒙古通辽028043 [2]内蒙古民族大学乳源性致病菌研究所,内蒙古通辽028043 [3]内蒙古民族大学动物科技学院,内蒙古通辽028043

出　　处：《中国病原生物学杂志》2012年第11期831-834,共4页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.31060325);内蒙古自治区科技厅科技创新引导计划项目(No.20111802;2012年)

摘　　要：目的克隆牛乳腺炎金黄葡萄球菌弹性纤维结合蛋白N端蛋白基因,并进行序列分析。方法根据Gen-Bank上公布的金黄葡萄球菌弹性纤维结合蛋白N端蛋白(nEBPS)基因的全序列,利用生物学软件Primer5.0和Oligo6.0设计了1对特异性引物,采用PCR方法扩增临床分离株nEBPS基因片段。结果扩增的基因片段长度为720bp,与标准菌株(CMCC26074)的nEBPS基因片段序列相似性为100%,与GenBank上公布的金黄葡萄球菌菌株(U48826.2)nEBPS基因相似性为97.36%,核苷酸序列共有19处出现碱基突变。结论成功克隆出牛乳腺炎金黄葡萄球菌SEB基因,为建立快速检测牛乳中金黄葡萄球菌的PCR检测方法奠定了基础。Objectives To clone and sequence the nEBPS gene for the N-terminal domain of the Staphylococcus aureus fibronectin-binding protein in cattle with mastitis.Methods In accordance with the sequence for the gene for the N-terminal domain of the S.aureus fibronectin-binding protein published on GenBank,a pair of primers was designed using Primer 5.0 and Oligo 6.0 software.nEBPS gene fragments from clinical isolates were amplified using PCR.Results Sequencing indicated that the nEBPS gene sequences were 720 bp in length.nEBPS gene sequences of clinical isolates had 100% similarity to gene sequences of the standard strain（CMCC26074）and had 97.36% similarity to the EBPS gene sequences of the standard S.aureus strain（U48826.2） published on GenBank.Nucleotide sequences had 19 base mutations.Conclusion The bovine mastitis S.aureus nEBPS gene was cloned and sequenced.This work has laid the foundation for rapid detection of S.aureus in milk.

关 键 词：金黄葡萄球菌 nEBPS基因 克隆 序列分析 

分 类 号：R378.11[医药卫生—病原生物学]