肝靶向肽-抗肿瘤肽CSP-MDA-7/IL-24融合基因原核表达条件优化  

Optimization of Expression Conditions of Induction Strategy for Improving Liver Targeting-Antitumor CSP-MDA-7/IL-24 Production in E.coli

作　　者：陈瑜丽 张晶晶 马艳 卢雪梅 刘文彬 金小宝 汪洁 CHEN Yu-Li;ZHANG Jing-Jing;MA Yan;LU Xue-Mei;LIU Wen-Bin;JIN Xiao-Bao;WANG Jie(School of Basic Sciences,Guangdong Pharmaceutical University,Guangzhou 510006;Guangdong Key Laboratory of Pharmaceutical Bioactive Substances,Guangzhou 510006;School of Biosciences and Biopharmaceutics,Guangdong Pharmaceutical University,Guangzhou 510006 China)

机构地区：[1]广东药科大学生命科学与生物制药学院,广东广州510006 [2]广东药科大学基础学院药用生物活性物质研究所,广东广州510006 [3]广东省生物活性药物研究重点实验室,广东广州510006

出　　处：《生物技术通讯》2018年第4期465-470,共6页Letters in Biotechnology

基　　金：广东省医学科学技术研究基金(A2016091)

摘　　要：目的:优化肝靶向肽-抗肿瘤肽(CSP-MDA-7/IL-24)在大肠杆菌中诱导表达的相关条件。方法:通过考察CSP-MDA-7/IL-24重组菌生长曲线,选择合适的诱导时机,通过SDS-PAGE检测CSP-MDA-7/IL-24蛋白的表达量,单因素分析诱导时间、培养基p H值、诱导温度和诱导剂IPTG浓度,在此基础上各选取5个水平进行正交实验,摸索最佳诱导表达条件。结果:培养基p H7.0、IPTG浓度0.12 mmol/L、培养温度42℃、诱导表达4 h为重组蛋白的最佳表达条件。结论:为进一步制备重组蛋白CSP-MDA-7/IL-24及评价其生物活性奠定了基础。Objective： To optimize the conditions for the induction of CSP-MDA-7/IL-24 expression in E.coli. Methods： Accoding to CSP-MDA-7/IL-24 recombinant strain growth curve, the appropriate timing of induction was determined. From SDS-PAGE of bacterial culture supernatant, the range of single factor induction time, medi- um pH value, induction temperature and IPTG inducer concentration were analyzed. Five levels of the four factors were selected for orthogonal test. Results： The results showed that induction temperature was the main factor influ- encing expression, pH8.0, IPTG concentration 0.12 mmol/L, 42℃, and 4 h induction for expression of recombinant protein. Conclusion： This study laid the foundation for the further preparation of recombinant protein CSP-MDA-7/ IL-24 and evaluation of its biological activity.

关 键 词：肝靶向肽-抗肿瘤肽 原核表达 正交试验 条件优化 

分 类 号：Q78[生物学—分子生物学]