肝靶向肽-抗肿瘤肽在大肠杆菌中诱导表达的相关条件优化研究  

Study on the optimization of the related conditions for the induced expression of liver targeting peptide-antitumor peptide in Escherichia coli

作　　者：单鹏[1] 刘现翠[2] SHAN Peng;LIU Xian-cui(Department of Pharmacy,Tengzhou Central People’s Hospital,Tengzhou 277599,China;Department of Pharmacy,Third People’s Hospital of Jinan,Jinan 250132,China)

机构地区：[1]滕州市中心人民医院药学部,滕州277599 [2]济南市第三人民医院药剂科,济南250132

出　　处：《药物分析杂志》2020年第9期1559-1564,共6页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:优化重组蛋白肝靶向肽-抗肿瘤肽(CSP-MDA-7/IL-24)在原核表达系统中的培养条件,并提高其蛋白的表达量。方法:对重组质粒在菌株BL21中的生长情况进行检查,确定合适的诱导时机;通过单因素试验,优化适合重组蛋白最大化表达的培养条件;并在此基础上设计4个因素5个检测水平的正交实验,从而确定最佳诱导重组蛋白表达的条件组合。结果:在pH为7,IPTG浓度为0.15 mmol·L^-1,温度为42℃、诱导时间为4 h时,重组蛋白表达百分比达到最大值。此外,分析显示在所研究的培养条件中,温度对重组蛋白表达水平的影响最为显著。结论:本研究结果为扩大重组蛋白CSP-MDA-7/IL-24的生产提供了数据支持,并为后期CSP-MDA-7/IL-24的生物学研究提供了研究材料。Objective:To optimize the cultural conditions for the expression of the recombinant protein CSP-MDA-7/IL-24 in prokaryotic system(E.coli)to upregulate the secretion of this target protein.Methods:By examining the expression level of recombinant plasmid in strain BL21,the proper growth curve was determined;Single factor test was used to optimize the appropriate culture conditions suitable for maximized expression of recombinant protein.Besides,orthogonal experiments of four factors with five levels for each were used to determine the optimum conditions for inducing recombinant protein expression of CSP-MDA-7/IL-24.Results:The secretion of recombinant protein expression reached the maximum level after induction of IPTG of 0.15 mmol/L at 42℃for 4 hours(pH 7.0).In addition,statistical analysis showed that temperature,instead of other examined parameters,exerted a significant effect on the expression of recombinant protein in the culture conditions mentioned above.Conclusion:This study provides the data support for the further expanding the production of recombinant protein CSP-MDA-7/IL-24 and its biological research.

关 键 词：肝靶向肽-抗肿瘤肽 原核表达系统 单因素实验 正交试验 优化培养条件 

分 类 号：R917[医药卫生—药物分析学]