增强型绿色荧光蛋白与IgG抗体亲和肽融合蛋白的制备  

Construction of the fusion protein between EGFP and IgG-binding peptide

作　　者：杨洪鸣[1] 张义军[1] 高鹏[1] 

机构地区：[1]潍坊医学院应用药理学研究室,山东潍坊261042

出　　处：《中国海洋药物》2007年第5期25-28,共4页Chinese Journal of Marine Drugs

摘　　要：目的构建、表达增强型绿色荧光蛋白(EGFP)与IgG抗体亲和肽融合蛋白,并对其生物学功能进行研究。方法将EGFP基因与IgG抗体亲和肽基因重组,构建原核分泌表达载体pSpA-EGFP-His,通过竞争ELISA和荧光光谱测定,考察其在大肠杆菌中表达产物SpA-EGFP融合蛋白的生物学活性。结果SpA-EG-FP融合蛋白的相对分子质量42kD,与理论值相近,荧光光谱与文献报道一致,且能与SpA-Peroxidase竞争结合反应体系中的兔IgG抗体。结论SpA-EGFP融合蛋白在E.coliDH5α中正确表达,且具有EGFP的荧光特性和与哺乳动物IgG抗体结合的生物学活性。Objective Making the fusion protein of IgG-binding peptide with enhanced green fluorescent protein （EGFP） and determining its bioactivity. Methods The enhanced green fluorescent protein （EGFP） gene was cloned into pEZZ 18 vector containing ZZ peptide gene to construct expression vector pSpA-EGFP-His. The fusion protein was expressed in E. coliDH5a and its bioactivity was detected by competitive ELISA and fluorescence properties. Results The fusion protein migrated at approximately 42kD in SDS-PAGE, which correspond to the theoretical molecular weight. The spectra of SpA-EGFP fusion protein was similar to what was reported. SpA-EGFP competed with SpA-Peroxidase to bind IgG. Conclusion The plasmid pSpA-EGFP-His correctly expressed in E. coll. The fusion protein retains the bifunctional effects of EGFP and IgG-binding activity.

关 键 词：增强型绿色荧光蛋白 IgG亲和肽 融合蛋白 

分 类 号：R915[医药卫生—微生物与生化药学]