葡萄球菌蛋白A“ZZ”与EGFP的融合表达及活性测定  

Expression and biological assay of ZZ-EGFP fusion protein in E. coli

作　　者：唐金宝[1] 宋淑亮[2] 吉爱国[2] 朱鹏[2] 

机构地区：[1]潍坊医学院基础医学部,261042 [2]山东大学威海生物技术研发中心,262049

出　　处：《中华微生物学和免疫学杂志》2007年第6期532-535,共4页Chinese Journal of Microbiology and Immunology

摘　　要：目的将EGFP基因与葡萄球菌蛋白A(SPA)基因的ZZ结构域重组,尝试以融合标记的方式构建一种新型免疫亲和试剂。方法利用基因工程技术将EGFP基因重组至pEZZ 18质粒,构建原核分泌表达载体pEZZ-EGFP,通过竞争ELISA和荧光光谱测定其在大肠杆菌中表达产物ProZZ- EGFP的生物学活性。结果pEZZ-EGFP在E.coli HB101中正确表达,所表达融合蛋白具有与哺乳动物IgG抗体结合的生物学活性和EGFP的荧光活性。结论ProZZ-EGFP融合蛋白有ZZ结构域(SPA)和EGFP二者生物学特性,在免疫荧光测定中可作为一种通用亲和试剂。Objective To express the fusion protein of enhanced green fluorescent protein （EGFP） with ＂ZZ＂ peptide of staphylococcal protein A （SPA） in E. coli and to test its bioactivity. Methods The fragment of EGFP gene was cloned into pEZZ 18 vector containing ZZ peptide gene. The recombinant plasmid was transferred into E. eoli HB101. The fusion protein was expressed in E. coli and its bioaetivity was examined by competitive ELISA and fluorescence properties. Results The plasmid pEZZ-EGFP correctly expressed in E. eoli. The fusion protein retains the bifunetional effects of ＂ZZ＂ peptide and EGFP. Conclusion The fusion protein of ZZ peptide with EGFP was expressed as a signal protein.ZZ peptide selectively binds to immunoglobulins G （IgG） of many mammalian, via Fe region of IgG. So the fusion protein could be applied to immunoassay, as a new alternative universal reagent.

关 键 词：ZZ结构域 增强型绿色荧光蛋白 融合蛋白 免疫测定 

分 类 号：Q93[生物学—微生物学]