SPG血清白蛋白结合区和RSV的G蛋白片段融合蛋白的构建和表达

Expression and purification of fusion protein containing the segment of glycoprotein G of RSV and the serum albumin-binding region of streptococcal protein G(BB)

目的构建表达呼吸道合胞病毒(RSV)G蛋白片段(来自130-230氨基酸)和来自链球菌蛋白G(SPG)血清白蛋白结合区(BB)的原核表达质粒,寻找适合的表达和纯化条件,获得融合表达蛋白,为进行体内外实验检测奠定基础.方法利用PCR技术,通过提取病毒cDNA和链球菌基因组DNA为模板扩增目的片段,构建融合表达质粒PET32a(BBG2Na)。转化宿主菌E. coli Bl21(DE3)进行表达,并采用亲和层析纯化目的蛋白。结果Thx-G2Na-BB在E. coli BL21(DE3)中可获得高效表达,表达量可达菌体蛋白的68.3%,并以可溶性的形式表达,纯化后的目的蛋白纯度可达90%。结论实现Thx-G2Na-BB的融合表达,此蛋白可作为抗原用于重组蛋白疫苗的筛选。

Objective To construct a prokaryotic expression system for the amino-acid region 130-230 of glycoprotein G and the serum albumin-binding region of SPG,and search an appropriate induction and purification conditions for gaining soluble fusion protein. Methods G2Na gene and BB gene were amplified by PCR with cDNA of RSV and DNA of streptococcal genome as templates,respectively. The recombinant expression plasmid PET32a (BBG2Na) containing Thx-BB-G2Na fusion gene was constructed by recombinant DNA technique,an...