J4 ›› 2010, Vol. 36 ›› Issue (2): 262-266.

• 基础研究 • 上一篇    下一篇

乳糖诱导角质细胞生长因子-2在大肠杆菌中的表达

田海山1,2,3|唐禄1,2|王晓杰1,3|王会岩1,2|刘笑菊1|冯秀萍1|王一1,2|李校堃1   

  1. 1.吉林农业大学 生物反应器与药物开发教育部工程研究中心|吉林 长春 130118;2.吉林农业大学生命科学学院|吉林 长春 130118;3.长春格鲁斯特生物科技有限公司|吉林 长春 130012
  • 收稿日期:2009-10-09 出版日期:2010-03-28 发布日期:2010-03-28
  • 通讯作者: 李校堃(Tel:0431-84533348,E-mail:xiaokunli@163.net) E-mail:xiaokunli@163.net
  • 作者简介:田海山(1972-)|男|吉林省长春市人|高级工程师|在读生物技术博士|主要从事生物反应器与基因工程药物研究。
  • 基金资助:

    国家863计划项目资助课题(2007AA02Z110);2009年度科技型中小企业技术创新基金项目资助课题(09C26212203306);吉林省长春市应用技术计划研究技术项目资助课题(08YZ45)

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

TIAN Hai-Shan1,2,3, TANG Lu1,2, WANG Xiao-Jie1,3, WANG Hui-Yan1,2, LIU Xiao-Ju1, FENG Xiu-Ping1, WANG Yi1,2, LI Xiao-Kun1   

  • Received:2009-10-09 Online:2010-03-28 Published:2010-03-28

摘要:

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10 g/L乳糖在33℃下诱导6 h,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。

关键词: 角质细胞生长因子-2;乳糖诱导;IPTG

Abstract:

Abstract: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 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.

Key words: keratinocyte growth factor -2;lactose induction;IPTG

中图分类号: 

  • Q78