猪链球菌2型新毒力相关基因lin0523的克隆表达及生物信息学分析  

Cloning,expression and bioinformatics analysis of a novel virulence-related gene lin0523 from Streptococcus suis serotype 2

作　　者：何永聚[1,2,3] 刘军[2,3] 李鹏[1,2,3] 孙洋[2,3] 祝令伟[2,3] 周博[2,3] 王文东[1] 冯书章[2,3] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]军事医学科学院军事兽医研究所,吉林长春130062 [3]吉林省人兽共患病预防与控制重点实验室,吉林长春130062

出　　处：《中国兽医学报》2011年第5期706-709,共4页Chinese Journal of Veterinary Science

基　　金：国家"973"计划资助项目(2006CB504402);吉林省科技发展计划资助项目(20080130)

摘　　要：lin0523是新鉴定出的猪链球菌2型(Streptococcus suisserotype 2,SS2)毒力相关基因。利用PCR方法扩增SS2野生强毒菌株ZY458的lin0523基因,并克隆到pMD18-T载体上。经限制性内切酶EcoRⅠ、PstⅠ双酶切后,将目的基因与大肠杆菌表达载体pMAL-p2x连接,构建重组表达载体pMAL-p2x::lin0523,并将其转化至宿主菌TB1中。经IPTG诱导后,表达出约为90 000的重组蛋白MBP-lin0523,目的蛋白主要以包涵体形式存在。应用同源性比对、信号肽预测、跨膜区预测及蛋白亚细胞定位等生物信息学方法对目的蛋白进行分析表明,lin0523蛋白与SS2 05ZYH33的限制性核酸内切酶S亚基100%同源,可能含有信号肽并存在于细胞外;这些结果表明lin0523蛋白可能是一种分泌性蛋白。lin0523 is a new identified virulence-related gene of Streptocuccus suis serotype 2.In this study,lin0523 was amplified from SS2 ZY458 isolated from Ziyang of Sichuan province,and cloned into pMD18-T.After the restriction enzyme digestion by EcoRⅠ and Pst Ⅰ,the lin0523 gene was subcloned into expression vector pMAL-p2x.The recombinant plasmid pMAL-p2x：lin0523 was transformed into E.coli TB1,then induced by IPTG at 37℃.The MBP-lin0523 was expressed in form of inclusion body,the recombinant protein molecular weight is about 90 000.The recombinant protein purified by amylose column.The characters of this target protein were analyzed by several online bioinformatics tools,including BLAST,SinglP,HMMTOP and PSORTb.The lin0523 protein may contain a signal peptide and a transmembrane region.It most likely present in the extracellular.These results showed that the lin0523 protein may be a secretory protein.

关 键 词：猪链球菌2型 lin0523基因 克隆表达 

分 类 号：S852.61[农业科学—基础兽医学] R535[农业科学—兽医学]