中国动物传染病学报2016，Vol.24Issue(2)：25-30,6.
2型猪链球菌SsU05-0474的表达纯化及单克隆抗体制备
EXPRESSION AND PURIFICATION OF SsU05-0474 OF STREPTOCOCCUS SUIS 05ZYH33 AND PREPARATION OF SPECIFIC MONOCLONAL ANTIBODY
杨宝玲 1陈福广 2谢芳 2刘思国 2沈国顺 1张跃灵2
作者信息
- 1. 沈阳农业大学畜牧兽医学院,沈阳110866
- 2. 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,哈尔滨150001
- 折叠
摘要
Abstract
To determine the location and formation of the major pilus subunit SsU05-0474 (SsMps) in Streptococcus suis serotype 2 (Ss2), the gene coding for SsMps was amplified in PCR from the genome of Streptococcus suisserotype 2 strain 05ZYH33 and cloned into pET22b to construct the expression plasmid pET22b/SsMps, which was then transformed into E. coli BL21(DE3). The recombinant SsMps was expressed with induction of IPTG and purified in affinity chromatography. BALB/c mice were immunized with purified SsMps. The anti-SsMps monoclonal antibody was prepared via cell fusion and subsequent screening. The results showed that the recombinant SsMps protein, with a molecular mass of about 45 kDa, was expressed as soluble form. About 30 mg of SsMps with purity above 95% could be obtained from 1 L of E. coliculture. One monoclonal antibody 1D9 was obtained and determined to be subtype of IgG1 withκ chain. As demonstrated in Western blot, the monoclonal antibody ID9 specifically reacted with natural SsMps in cellular wall fraction of 05ZYH33 and SsMps was presentas polymers with different lengths. The availability of purified SsMps protein and monoclonal antibody laid solid foundation for further research on its functions.关键词
2型猪链球菌/SsMps基因/蛋白表达/蛋白纯化/单克隆抗体Key words
Streptococcus suisserotype 2/SsMps gene/protein expression/protein purification/monoclonal antibody分类
农业科技引用本文复制引用
杨宝玲,陈福广,谢芳,刘思国,沈国顺,张跃灵..2型猪链球菌SsU05-0474的表达纯化及单克隆抗体制备[J].中国动物传染病学报,2016,24(2):25-30,6.
