hcv ns5b蛋白对hcv rna 的模板特异性研究3.pdf

HCV NS5 B 蛋白对 HCV RNA 的模板特异性研究 叶　力 ,叶林柏 ,佘应龙 , Khalid A mine Timani ,廖庆姣 ,吴正辉 , 郜金荣 ,孔令保 ,李宝宗 , 曾莹春 (武汉大学生命科学学院病毒学国家重点实验室 ,湖北武汉 ,430072) Template Specif icity of HCV NS5 B in RNA Syntheses f rom 3 ′Terminal Sequences of HCV Positive and Negative Strand RNA YE Li , YE Linbai , SH E Yinglong , Khalid A mine Timani , L IA O Qingj iao , WU Zhenghui , GA O J inrong , KON G Lingbao , L I Baozong , ZEN G Yinchun ( ) S t ate Key L aboratory of Vi rology , Col leg e of L if e S ciences , Wuhan Uni vers ity , Wuhan 430072 , Chi na Abstract : A reco mbinant H CV RN Adep endent RN A polymera se ( RdRp , N S5B p rot ein) wa s ex ( ) p re ssed in E. col i st rain BL 2 1 D E3 . The p urified p rot ein wa s u sed to inve stigat e t he RN A syn t he se s f ro m t he 3′t er minal sequence s of H CV po sitive and negative st rand RN A i n v i t ro . The po sitive st ran d RN A did not direct t he synt he si s of negative st rand RN A ; however , t he negative st rand RN A wa s able to generat e a f ulllengt h p o sitive st ran d RN A . The se re sult s demon st rat e t hat N S5B ha s t he t emp lat e sp ecificit y on negative st rand RN A . The t emp lat e sp ecificit y wa s f ur t her confir med by t emp lat e co mp etition a ssay . The co mp etition of po sitivest rand RN A did not aff ect t he RN A synt he si s f ro m t he negative st rand RN A .