GST融合的人血浆蛋白S成熟肽的原核表达  

PROKARYOTIC EXPRESSION OF GST- FUSD MATURE PEPTIDE OF HUMAN PROTEIN S

作　　者：卢爱薇[1] 兰风华[2] 程烽[2] 郑德柱[2] 谢飞[2] 朱忠勇[2] 

机构地区：[1]三明市第一医院检验科 [2]福州总医院实验科

出　　处：《福州总医院学报》2004年第1期18-20,共3页Journal of Fuzhou General Hospital

摘　　要：目的:构建人蛋白S(protein S,PS)的原核表达载体并诱导其表达。方法:自行设计引物,通过PCR技术,以人PS真核表达载体为模板扩增PS成熟肽编码序列,PCR产物经EcoR1/BamH1双酶切后,将其克隆至GST融合表达载体pGEX-2T中,在E.coli BL21中诱导GST-PS融合蛋白的表达。结果:聚丙烯酰胺凝胶电泳(10%SDS-PAEG) 显示,在IPTG的诱导下,BL21重组菌高效表达出一个分子量约为96kD产物,与预期大小相符,该产物可通过GST亲和层析柱纯化。对重组质粒的序列分析表明,插入片段的序列与Genebank登录的人PS基因编码序列完全一致。结论:人PS 编码序列已被克隆至GST融合表达载体pGEX-2T,并在E.coli BL21中获得表达,为进一步研究奠定了基础。Objective: To construct a prokaryotic vector for the expression of human protein s. Method: the coding sequence of PS mature peptide was amplified from a existing recumbinant-plasmid (PS pcDNA3) with primers designed in our laboratory,and cloned into pGEX - 2T after restriction . GST - PS fusion protein was expressed after induction with IPTG. Results: Sequencing and restriction digestion of the recombinant plasmid revealed the existence of coding sequence for PS mature peptide. SDS * PAGE showed that a protein hand of about 96KD could be induced by IPTG in the lysate of bacteria harboring the recombinant plasmid.Conclusion: The coding sequence of PS mature peptide was introduced into the pGEX - 2T plasmid and a GST - fused PS mature peptide could be induced at a high level.

关 键 词：GST 蛋白S 人血浆 成熟肽 诱导 融合表达载体 PS E.coli 亲和层析柱 原核表达 

分 类 号：R392[医药卫生—免疫学] R735[医药卫生—基础医学]