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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 construct prokaryotic expression vector of human annexin and induce its expression. Methods The coding sequence of annexin gene was amplified by RT-PCR from total RNA of human placenta tissue. The amplified product was cloned into GST fusion expression vector pGEX-2T, and the expression of GST fusion human annexin was induced by IPTG. Results The results of gel electrophoresis showed that the size of RT-PCR product in human placenta was consistent with the size of the target fragment. Analysis of the recombinant plasmids showed that the sequence of the inserted fragment was identical to that of the published human annexin gene. Under the induction of IPTG, the recombinant BL2 1 efficiently expressed a product with a molecular weight of about 61 kDa. Conclusions Human annexin coding sequence has been cloned into GST fusion expression vector pGEX-2T.