重组人a降钙素基因相关肽在大肠杆菌中表达条件的优化.pdf

7 4 生命科学研究 Vol.7 No.4 200312 Life Science Research Dec. 2003 A ¹ 赵丹丹, 于晓光, 班 莺, 邹海峰,林 平, 徐华锋 ( , 150086) : 为提高人A 降钙素基因相关肽( hACGRP) 在大肠杆 中的表达量,研究了本室构建的pET-hACGRP 重 组质粒在E . coli BL21trxB(DE3)pLysS 宿主 中的表达条件. 经SDS分析和 LN2000 凝胶影像分析结果 表明,重组 以LB为培养基, 氨苄青霉素浓度为50 mg/L, 开始诱导时的 体密度为 OD = 0.6 ~ 0.8, 所加 600 IPTG 的浓度为0.75 mmol/L,37 e 摇床180 ? 5 r/min 振荡诱导培养4 h 时, 可获得高效表达的hACGRP 融合蛋 白. 重组蛋白的最高表达量占 体总蛋白的70 % ~ 80 %; 它主要以可溶性形式存在, 为下 一步纯化工作提 供了方便. : 人A 降钙素基因相关肽;表达条件; 优化;SDS-聚丙烯酰胺凝胶电泳 : Q786 : A : 1007-7847(2003)04-0315-05 Optimization of Recombinant Human A Calcitonin Gene-related Peptide Expression in E . coli ZHAO Dan-dan, U Xiao-guang, BAN ying, ZOU Ha-i feng, LIN Ping, XU Hua-feng (Department of Biochemistry and Molecular Biology , Harbin Medical University, Harbin 150086, Heilongj iang, hina) Abstract: In order to improve the expression level of human A calcitonin gene-related peptide ( hACGRP) , the growth conditions of the engineering strain, that confirmed by transforming the recombinant plasmid pET-hACGRP which constructed by our team into the host E. coli BL21trxB(DE3) pLysS were studied,which remarkably influ- enced the final yield of protein expression. With the aid of SDSand LN2000 gel image analysis, it was found that the best express