摘要
目的 构建人膜联蛋白 的原核表达载体并诱导其表达。方法 利用 RT- PCR技术从人胎盘组织总RNA扩增膜联蛋白 基因的编码序列 ,将扩增产物克隆至 GST融合表达载体 p GEX- 2 T,以 IPTG诱导 GST融合人膜联蛋白 的表达。结果 凝胶电泳显示人胎盘组织 RT- PCR扩增产物的分子量大小与目的片段大小相符。对重组质粒分析表明 ,插入片段的序列与发表的人膜联蛋白 基因编码序列一致。在 IPTG的诱导下 ,BL2 1重组菌高效表达出一个分子量约为 6 1k Da产物。结论 人膜联蛋白 编码序列已被克隆至 GST融合表达载体 p GEX- 2 T。
Objective To express human annexin Ⅴ in E.coli .Methods The coding sequence of human annexin Ⅴ gene was amplified from human placenta total RNA by RT PCR.The PCR product was cloned into the GST fusion expression vector pGEX 2T and the fusion protein was induced by IPTG.Results A specific band of 980bp from the RT PCR amplification was seen in gel electrophoresis.A fragment of the same size could be amplified directly from the bacterial cells(BL21)transformed with the recombinant plasmid.The sequence of the insert in the plasmid was identical to the published coding region sequence of human annexin Ⅴgene.A protein product with a molecular weight of about 61KDa could be induced by IPTG.Conclusion The coding sequence of human annexin Ⅴ geng was successfully cloned into the GST fusion expression vector pGEX 2T.
出处
《福建医药杂志》
CAS
2002年第6期123-125,共3页
Fujian Medical Journal
关键词
鉴定
人膜联蛋白V
分子克隆
原核表达
Human annexin Ⅴ
Molecular cloning
Prokaryotic expression
