乳糖诱导mIL-9蛋白高效表达研究  

Research on High-level Expression of Lactose-induced Recombinant mIL-9

作　　者：王建洲[1,2] 罗竞红[3] 彭红卫 周裕程 邓洪新[1] 

机构地区：[1]四川大学生物治疗国家重点实验室,四川成都610041 [2]成都金凯生物技术有限公司,四川成都610041 [3]成都大学城乡建设学院,四川成都610106 [4]诺迪康药业股份有限公司,四川成都610016

出　　处：《成都大学学报（自然科学版）》2014年第2期106-109,129,共5页Journal of Chengdu University（Natural Science Edition）

摘　　要：研究了乳糖诱导重组工程菌pET(32a+)-rmIL-9/BL21(DE3)高效表达rmIL-9融合蛋白的实验条件,探讨规模化生产rmIL-9的可行性.分别考察乳糖诱导时机、诱导浓度、诱导时间等参数对rmIL-9融合蛋白表达的影响,并通过正交实验,筛选最佳的乳糖诱导条件.结果显示,对于rmIL-9融合蛋白,乳糖作为诱导剂可达到良好的诱导效果,诱导产物的表达量可优于IPTG诱导产物.最优诱导表达条件为:当菌液OD600值为1时,添加终浓度为5g/L的乳糖,诱导9 h可以高效表达目的蛋白.实验表明,采用乳糖可以诱导rmIL-9融合蛋白基因的高效表达,此为动物实验评价rmIL-9治疗黑色素瘤提供了前期基础.To study the experimental conditions of high-level expression of rmIL- 9 in E. coli BL21 （DE3） with lactose instead of IPTG and explore the feasibility of rmIL - 9 scale production, we investigate respec- Lively the effects of main parameters such as lactose induction timing, the inducing concentration and the duration of induction on the fusion protein expression of rmlL - 9, and then screen the optimal lactose in- duction conditions by orthogonal test. The experimental results show that for nnlL- 9 fusion proteins, lac- tose as an inducer can achieve good induction effect, and the amount of expression of inducted product can be superior to that of inducted product by IPTG. The optimal induction expression condition is that when the bacterial concentration reaches OD600 = 1 and the lactose of final concentration 5 g/L is added, the target proteins can be highly expressed after being induced for 9 hours. We conclude that lactose can be used as a promising inducer in the production of recombinant mIL - 9, which provides a preliminary basis for the treatment evaluation of rmIL- 9 in melanoma.

关 键 词：重组mIL-9 乳糖 高效表达 

分 类 号：R914.2[医药卫生—药物化学]