乳糖诱导角质细胞生长因子-2在大肠杆菌中的表达

Expression of KGF-2 in E. coli induced with lactose

目的:研究用乳糖替代IPTG作为诱导剂进行角质细胞生长因子-2(KGF-2)基因重组蛋白的表达,探讨工程化生产的可能性。方法:分别比较乳糖诱导浓度、诱导时机、诱导时间及诱导温度对重组表达产物的影响,并以IPTG诱导作为对照,确定较为优化的乳糖诱导条件。结果:对于重组目的产物,乳糖作为诱导剂也可以起到良好效果,以乳糖为诱导剂的A600值较IPTG高,诱导产物的表达量与IPTG诱导产物相当。以终浓度为10g.L-1乳糖在33℃下诱导6h,可以达到良好的诱导效果,并且具有较好的可溶性和促细胞生长活性。结论:乳糖可替代IPTG诱导角质细胞生长因子基因表达,并取得良好效果,为工程化生产提供了良好的实验依据。

Objective To study the expression of the recombinant protein of keratinocyte growth factor-2（KGF-2） induced with lactose,an alternative to IPTG induction agents,and explore the possibility of the production of KGF-2 engineering.Methods The effects of the concentration,induction time and induction temperature of lactose on the expression of the recombinant product were compared and IPTG induction was used as a control to determine a more optimal conditions for lactose induction.Results For the production of recombinant KGF-2,lactose as an induction agent could also play a very good role;the A600 value with lactose as induction agent was higher than IPTG,the expression level of induction product was equivanlent to IPTG induction.With 10 g·L^-1 lactose,at 33℃,induction for 6 h,the induction effect was better and the induction product had good solubility and could promote the cell growth.Conclusion Lactose can replace IPTG to express the keratinocyte growth factor,and the good effects are obtained.This study provides a good experimental basis for the large production of KGF-2.