SHV-12型超广谱β-内酰胺酶的原核表达及免疫原性分析  

Prokaryotic Expression of SHV-12 Type ESBLs and Immunogenicity Analysis

作　　者：贾芳[1,2] 周学章[1,2] 

机构地区：[1]宁夏大学生命科学学院,宁夏银川750021 [2]宁夏大学西部特色生物资源保护与利用教育部重点实验室,宁夏银川750021

出　　处：《宁夏大学学报（自然科学版）》2011年第2期164-167,共4页Journal of Ningxia University(Natural Science Edition)

基　　金：国家自然科学基金资助项目(31060348);宁夏科技攻关基金资助项目(KGZ-12-09-06)

摘　　要：利用PCR技术扩增了SHV-12 ESBLs目的基因,并将目的基因亚克隆到表达载体pET-28a(+)中,获得重组质粒pET-28a-SHV-12转化于表达菌株BL21(DE3)中诱导表达,表达蛋白纯化后经SDS-PAGE电泳分析.结果表明,目的蛋白SHV-12 ESBLs相对分子质量为30 KDa,经蛋白质印迹检测证明具有特异性条带,经头孢硝基噻吩显色反应表明获得了有活力的ESBLs,包涵体作为免疫抗原免疫Balb/c小鼠,3次免疫后利用间接ELISA法测定免疫效价达到1∶106.获得了可靠而稳定的免疫原.SHV-12 ESBLs target gene was amplified by PCR and subcloned into a vector pET-28a（＋）.The positive plasmid contained SHV-12 ESBLs gene was transferred into BL21（DE3） and was induced to expression and purification by IPTG.It proved that the SHV-12 ESBLs was expressed and purified as a molecular weight 30 KDa fusion protein which was verified by SDS-PAGE.Western blotting of the results showed that there was specific band.Nitrocefin colour reaction certificated that there was dynamic ESBLs.Inclusion body can stimulate Balb/c mice to produce high titer antibodies to 1∶106,which the recombinant ESBLs can be used as immune with good immunogenicity.

关 键 词：克隆与表达 超广谱β-内酰胺酶(ESBLs) 抗体效价 

分 类 号：Q556[生物学—生物化学]