OCT蛋白对猪链球菌生物被膜形成和细胞黏附的影响  

Effect of OCT protein on the biofilm formation and the bacterial adherence of Streptococcus suis

作　　者：梅京京 汪洋[1,2] 易力[3] 张建康[1] 程相朝[1] 张春杰[1] 余祖华[1] 吴庭才[1] 

机构地区：[1]河南科技大学动物疫病与公共安全重点实验室,河南洛阳471003 [2]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,甘肃兰州730046 [3]洛阳师范学院生命科学学院,河南洛阳471022

出　　处：《中国兽医科学》2016年第2期192-197,共6页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31201910;31540095);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)开放课题(SKLVEB2013KFKT003);河南省高校创新人才基金项目(14HASTIT024)

摘　　要：根据GenBank中猪链球菌鸟氨酸氨甲酰转移酶基因(oct)序列(GenBank登录号:NZ_CP007497)设计引物,克隆猪链球菌ZY05719株的oct基因,构建p ET-28a-oct原核表达质粒并在大肠杆菌Rosetta中诱导表达,重组OCT蛋白即鸟氨酸氨甲酰转移酶用镍亲和层析柱纯化后,研究它对猪链球菌生物被膜形成的影响及介导猪链球菌黏附PK-15细胞的作用。结果显示,克隆出1 014 bp的oct基因,并检测出纯化的40 ku OCT融合蛋白具有良好的反应原性,且能提高猪链球菌形成生物被膜的能力及降低黏附PK-15细胞的作用。本研究成功克隆并表达了ZY05719株的oct基因,初步揭示了其生物学功能,为深入研究鸟氨酸氨甲酰转移酶在猪链球菌致病机制中的作用奠定了基础。The oct gene fragment was amplified by PCR from Streptococcus suis ZY05719 strain and cloned into prokaryotic expression plasmid p ET-28 a to form p ET-28a-oct. The expression of recombinant protein was induced in E.coli Rosetta and was purified. The expressed and purified proteins were analyzed by SDS-PAGE and its antiserum was raised from immunized rabbits. The effect of purified recombinant protein and its antiserum on the biofilm formation and on adhesion of S.suis to PK-15 were assayed.In result, sequence analysis showed that the length of the oct gene was 1 014 bp. The prokaryotic expressed product was a fusion protein, whose molecular weight was about 40 ku and could improve the S.suis biofilm formation and could keep down adhesion of S.suis to PK-15. The oct gene of ZY05719 has been successfully cloned and highly expressed in E.coli Rosetta, the recombinant protein facilitates the further studies on the biofilm and the bacterial adhesion, and even the pathogenesis of S. suis.

关 键 词：猪链球菌 鸟氨酸氨甲酰转移酶 生物被膜 细胞黏附 

分 类 号：S852.611[农业科学—基础兽医学]