重组人乙型肝炎病毒核糖核酸酶H基因工程菌发酵条件的初步研究  

Preliminary studies on fermentation of the recombinant human HBV-RNase H in E.coli

作　　者：邹爱民[1] 高萍[2] 林芳[1] 张利朝[1] 张惠中[1] 

机构地区：[1]第四军医大学中心实验室,陕西西安710038 [2]第四军医大学妇产科,陕西西安710038

出　　处：《中国生化药物杂志》2005年第2期80-83,共4页Chinese Journal of Biochemical Pharmaceutics

摘　　要：目的研究表达乙型肝炎病毒核糖核酸酶H1及H2 (HBV -RNaseH1及H2 )工程菌的发酵条件。方法采用摇瓶发酵,研究不同宿主菌表达HBV- RNaseH1及H2的效果,同时对培养基、诱导时期、诱导时间、初始pH值等发酵条件以及质粒稳定性进行研究。结果选用EcoliBL2 1为宿主菌,在LB培养基中培养至A60 0nm为0 .6时,诱导表达4 .5h ,HBV- RNaseH1及H2表达量最高达33.6 %和30 .8%。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较好地提高HBV- RNaseH1及H2的表达量。PurposeTo study the fermentation process of recombinant bacteria E.coli BL21 expressing recombinant human HBV-RNase H gene.MethodsThe E.coli host cells, culturemedium, induction period, induction time and pH were optimized. The stability of pGSTag/HBV-RNase H1 and pGSTag/HBV-RNase H2 plasmid was also investigated. ResultsThe expression level of HBV-RNase H1 was about 33.6% and H2 was about 30.8% in the total bacteria protein, when recombinant E .coli BL21 harboring pGSTag/ HBV-RNase H1 and H2 plasmid was induced for 4.5 hours after the bacteria density A_(600nm) reached 0.6 in LB medium. The pGSTag/HBV-RNase H1 and H2 plasmind in E.coli BL21 remained stable, after being cultured to 60 generations.ConclusionThe developed fermentation technique might increase the expression level of HBV-RNase H1 and H2,and is suitable for the fermentation of HBV-RNAse H1 and H2 on a large scale.

关 键 词：基因工程菌 人乙型肝炎病毒 核糖核酸酶H基因 发酵 

分 类 号：R373.21[医药卫生—病原生物学]