小鼠OX40原核表达载体的构建及表达  

Construction of the Prokaryotic Expression Vector of Mouse OX40 Gene and Its Expression

作　　者：林志娟[1] 刘艳菲[1] 冯永堂[1] 许传武[1] 杜雪梅[1] 苗乃法[1] 

机构地区：[1]潍坊医学院免疫学教研室山东省高校免疫学重点实验室,山东潍坊261042

出　　处：《潍坊医学院学报》2008年第3期202-204,共3页Acta Academiae Medicinae Weifang

摘　　要：目的构建小鼠OX40胞外段基因的原核表达载体并进行诱导表达。方法PCR扩增OX40胞外段基因并将其插入原核表达质粒pET32a(+),重组pET32a-OX40经测序鉴定后转入表达菌株BL21(DE3)进行融合蛋白的小量诱导表达。结果出现新增蛋白条带,融合蛋白主要存在于包涵体中;Western Blotting分析表明该融合蛋白可与相应抗体发生特异性结合。结论构建了小鼠OX40基因的原核表达载体,获得OX40胞外段融合蛋白。Objective To establish a prokaryotic expression vector for mouse OX40 extracellular gene,and induce it to express the protein in Ecoli. BL21（ DE3 ）. Methods The mouse OX40 extracellular gene was amplified by PCR and inserted into pET32a（ ＋ ）. The combinant vector pET32a-OX40 was analyzed by DNA sequencing,PCR and enzyme digestion. After induction with IPTG,SDS-PAGE and Western Blotting was applied to identify whether we have got the product of our protein. Results DNA sequencing indicated it was consistent with OX40 extracellular gene reported in GenBank. SDS-PAGE showed a new protein in pET32a-OX40/DE3 and the protein was mainly exsisting in inclusion body of bacteria. Western blotting analysis suggested that the new protein could bind specifically with anti-His · tag antibody. Conclusion We have got the fusion protein of extracellular OX40 by establishing pET32a-OX40.

关 键 词：OX40 载体构建 原核表达 

分 类 号：R392.12[医药卫生—免疫学]