最小弧菌热不稳定型溶血素表达载体的构建和高效表达  

Constrcuction and high expression of a vmha vector of Vibrio mimicus

作　　者：李涛[1] 吴后波[1] 

机构地区：[1]中国科学院南海海洋研究所热带海洋环境动力学重点实验室,广东广州510301

出　　处：《台湾海峡》2007年第2期242-248,共7页Journal of Oceanography In Taiwan Strait

基　　金："十五"国家高技术研究发展计划(863计划)资助项目(2003AA622050);国家自然科学基金资助项目(30371106)

摘　　要：根据已发表的最小弧菌热不稳定型溶血素(Vibrio mimicusheat-labile hemolysinVmhA)核苷酸序列,设计和合成一对特异性引物,以最小弧菌96-6-3的基因组DNA为模板,PCR扩增得到热不稳定型溶血素基因片段,克隆于表达载体PQE-2/VmhA中,转化大肠杆菌M15,经IPTG诱导,表达出预期大小50.2KD的融合蛋白.Westernblot检测显示,该蛋白与抗最小弧菌热不稳定型溶血素兔血清呈阳性反应,表明该蛋白具有溶血素的免疫原性,可作为基因工程疫苗的候选抗原.A pair of primers was designed according to the reported nucleotide sequences of the heatlabile hemolysin gene of Vibrio mimicus. With the specific primers, one target fragment was ampli- fied from Vibrio mimicus strain 96-6-3 DNA via PCR, the target fragment was cloned into an expression vector,PQE-2/VmhA. The recombiant protein exhibited a molecular weight of about 50.2KD in 10% SDS-PAGE, which was identical to what had been anticipated. Western blot analysis shows that rabbit polyantibody reacts to the recombiant protein, which indicated that the recombiant protein may have similar epitopes to the natural protein.

关 键 词：最小弧菌 溶血素 基因表达 

分 类 号：Q941.42[生物学—植物学]