人epiregulin在大肠杆菌中的表达  

Expression of Human Epiregulin in E.coli

作　　者：席全胜[1] 钱欣国[1] 周庆玮[1] 潘巍[1] 李载平[1] 甘人宝[1] 

机构地区：[1]中国科学院上海生物化学研究所,上海200031

出　　处：《生物化学与生物物理学报》2000年第3期295-298,共4页

摘　　要：利用RT PCR的方法首次从人肺癌细胞A5 49中扩增得到了epiregulin的cDNA。在 3′端添加 6个组氨酸密码子后将其克隆到本实验室构建的大肠杆菌高效碱性磷酸酶分泌表达系统中进行分泌表达。表达产物经Ni柱一步分离后得到纯化。对其进行氨基酸序列分析 ,结果与报道的人epiregulin的序列一致。MTT法测定其对Balb/c3T3细胞有很强的促增殖作用 ,而对表皮癌细胞A431有较强的抑制生长作用。Human epiregulin cDNA was amplified from the lung cancer cell line A549 using RT PCR. After adding 6 His codon to its 3′end, it was cloned into a high efficient secretive Escherichia coli system with alkaline phosphatase promoter( phoA promoter)constructed in our lab and induced for expression. The product was purified one step by Ni NTA column. Amino acid sequence analysis revealed the identity of our product with that previously reported. The product showed strong proliferative effect on fibroblast cell line Balb/c3T3 and growth inhibitory effect on epithelial carcinoma cell line A431.

关 键 词：EPIREGULIN 大肠杆菌表达系统 EGF 重组蛋白 

分 类 号：Q786[生物学—分子生物学]