乳糖诱导重组人睫状神经营养因子在大肠杆菌中的可溶性表达  

Soluble Expression of Recombinant Human Ciliary Neurotrophic Factor in E.coli Using Lactose as Inducer

作　　者：余永恒[1] 李清雄 王革非[1] 邓远鹏 王贞慧[1] 朱俊铭[1] 

机构地区：[1]丝宝集团生命科学研究所,武汉430058

出　　处：《中国生物制品学杂志》2007年第1期43-46,共4页Chinese Journal of Biologicals

摘　　要：目的利用乳糖替代IPTG,诱导重组人睫状神经营养因子(Recombinanthumanciliaryneurotrophicfactor,rhC-NTF)在原核工程菌BL-TCS中可溶性表达,并选择最适的诱导表达条件。方法在不同的温度下,利用不同浓度的乳糖及0·5mmol/L的IPTG,诱导含有人CNTF基因的工程菌9h,比较菌体生长、乳糖消耗以及目标蛋白的表达规律。结果乳糖组的菌体A600值均高于IPTG组。乳糖诱导产生的rhCNTF主要以可溶形式表达,各浓度乳糖的诱导效果均能接近或超过IPTG的效果。以2%乳糖在25℃下诱导6h,可以达到比较好的诱导效果。结论乳糖可取代IPTG诱导人CNTF基因表达,并获得良好效果。Objective To express recombinant human ciliary neurotrophic factor（rhCNTF） in E. coli BL-TCS,in a soluble form,using lactose as inducer instead of IPTG,and optimize the condition for expression. Methods Induce the expression of rhCNTF in E. coli BL-TCS with various concentrations of lactose at different temperatures for 9 h,using 0. 5 mmol/L IPTG as control. Compare the growth of bacteria, the consummation of lactose and the expression of target protein. Results All the A600 values of bacteria induced by lactose were signifieantly higher than that by IPTG and increased with the increasing lactose concentration. The rhCNTF under induction of lactose was mainly expressed in a soluble form ,and the expression level was not less than that under induction of IPTG. The condition for expression was optimized as incubation at 25℃ for 6 h under induction of 2% lactose. Conclusion Lactose might be used as an inducer of expression of rhCNTF in E. coli instead of IPTG.

关 键 词：乳糖 诱导剂 人睫状神经营养因子 大肠杆菌 

分 类 号：Q786[生物学—分子生物学]