重组雪花莲外源凝集素(GNA)表达条件的优化  

Optimization of Recombinant Galanthus Nixalis Agglutinin(GNA) Expression in E.coli

作　　者：罗素兰[1] 长孙东亭[1] 张真[1] 陈琴[1] 

机构地区：[1]海南大学生物技术中心海南大学海洋学院制药工程系,海南海口570228

出　　处：《药物生物技术》2005年第5期298-302,共5页Pharmaceutical Biotechnology

基　　金：国家自然科学基金资助项目(No:30260066);海南省自然科学基金资助项目(30111)

摘　　要：研究了含有GNA基因的重组大肠杆菌菌株G2和G3在不同诱导培养温度、起始诱导菌体密度(OD600值)、诱导剂异丙基-β-D-硫代半乳糖苷(IPTG)浓度及诱导培养时间等重要因子对重组GNA表达的影响,以期获得重组GNA表达的最佳条件.实验结果表明,不含信号肽的G2菌株和含有信号肽的G3菌株的最佳培养条件分别为:起始诱导菌体密度为OD600≈0.6;诱导剂IPTG浓度为0.1mmol/L;诱导培养时间为6 h;G2和G3的诱导培养温度分别为37℃,28～30℃.该研究结果将为发酵生产重组GNA的工艺提供依据,也为重组GNA开发成为生物农药、试剂和免疫增强剂提供支持.The effects of different induced culture temperatures and OD600 of initiated bacteria density on GNA expression in recombinant E. coli strain G2 and G3, as well as concentration of inducer IPTG and inducing cultivated time were studied in this research to optimize these parameters. The results showed that the optimized parameters of recombinant snowdrop lectin are as follows, the induced starting bacteria densities （OD600） were 0. 6, the IPTG concentrations were 0. 1 mmol/L, the best culture time was 6 h, the induced temperature of strain G2 was 37℃, and that of strain G3 with signal peptide was 28～ 30℃. The optimized conditions of recombinant GNA expression would increase the GNA production and decrease the cost. This research would be the basis for GNA fermentation production, and would use GNA as pesticide, reagent and immunity enhancer.

关 键 词：重组雪花莲外源凝集素 温度 诱导剂异丙基-β-D-硫代半乳糖苷 大肠杆菌表达 

分 类 号：Q81[生物学—生物工程]