C. He�, 王淑静, 戚基萍, 庄敏, 李冀宏, 徐建咏
Abstract
目的 建立成纤维细胞激活蛋白(FAP)表达系统,以获得完整蛋白.方法采用RT-PCR方法,自人新鲜胃癌组织中扩增FAP-cDNA,同时将FAP-cDNA克隆到pMD18-T中,表达正确后,再定向插入真核表达载体pQE30中,转化大肠埃希菌JM109,经异丙基-B-D-硫代半乳糖苷(IPTG)诱导表达.结果测定FAP-cDNA序列与GeneBank(基因库)中的FAP序列相同;构建出重组表达载体pQE30-FAP;转化大肠埃希菌JM109,并获得完整蛋白,经验证为诱导后的表达蛋白.结论成功构建pQE30-FAP表达载体,Westem-Blotting验证为完整蛋白,为进一步获得抗FAP抗体奠定了基础。
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目的 建立成纤维细胞激活蛋白(FAP)表达系统,以获得完整蛋白.方法采用RT-PCR方法,自人新鲜胃癌组织中扩增FAP-cDNA,同时将FAP-cDNA克隆到pMD18-T中,表达正确后,再定向插入真核表达载体pQE30中,转化大肠埃希菌JM109,经异丙基-B-D-硫代半乳糖苷(IPTG)诱导表达.结果测定FAP-cDNA序列与GeneBank(基因库)中的FAP序列相同;构建出重组表达载体pQE30-FAP;转化大肠埃希菌JM109,并获得完整蛋白,经验证为诱导后的表达蛋白.结论成功构建pQE30-FAP表达载体,Westem-Blotting验证为完整蛋白,为进一步获得抗FAP抗体奠定了基础。
Key concepts: Expression (computer science), Protein expression, Cell biology, Vector (molecular biology), Expression vector, Fibroblast, Biology, Computer science