摘要
目的:研究表达乙型肝炎病毒核糖核酸酶H(HBV RNaseH)工程菌的发酵条件。方法:采用摇瓶发酵,研究不同宿主菌对表达HBV RNaseH的效果;同时对培养基、诱导时期、诱导时间、初始 pH等发酵条件以及质粒稳定性进行研究。结果:选用E coliBL21为宿主菌,在LB培养基中培养至A600为0. 6时,诱导表达4. 5 h,HBV RNaseH表达量最高达33. 6%。而且重组质粒具有良好的分离稳定性和结构稳定性。结论:此发酵条件可以比较好的提高 HBV RNaseH的表达量,为发酵规模的扩大奠定了基础。
Objective: To study the optimization ferm entation conditions of recombinant genetic engineering bacteria expressing recom binant human HBV-RNaseH gene. Methods: The expressions of HB V-RnaseH from the different E.coli host cells were compared. The fermentation co nditions including culture medium, induction period, induction time and pH were optimized and the stability of pGSTag/HBV-RNAseH plasmid was investigated. Results: E. coliBL21 was selected as the host bacteria. The expre ssion level of HBV-RNaseH was about 33.6℅ in the total bacteria protein when re combinant E. coli BL21 harboring pGSTag/HBV-RNAseH plasmid was induced for 4.5 h ours after the bacteria density A600 reached 0.6 within LB medium. The pGSTag/HB V-RNAseH plasmind in E coli BL21 still remained stable after transfer of culture to 60 generations. Conclusion: The established fermentation conditions might increase the expression level of HBV-RNAseH and be suitable fo r the fermentation of HBV-RNAseH on a larger scale.
出处
《西南国防医药》
CAS
2005年第1期14-17,共4页
Medical Journal of National Defending Forces in Southwest China
