Expression and characterization of RNA-dependent RNA polymerase of Ectropis obliqua virus

被引：2

作者：

Lin, Meijuan ^{[1
]}

Ye, Shan ^{[1
]}

Xiong, Yi ^{[1
]}

Cai, Dawei ^{[1
]}

Zhang, Jiamin ^{[1
]}

Hu, Yuanyang ^{[1
]}

机构：

[1] Wuhan Univ, State Key Lab Virol, Coll Life Sci, Wuhan 430072, Hubei Province, Peoples R China

来源：

BMB REPORTS | 2010年 / 43卷 / 04期

基金：

中国国家自然科学基金;

关键词：

Ectropis obliqua virus; Iflavirus; Primer-dependent; RNA-dependent RNA polymerase; RNA synthesis; PICORNA-LIKE-VIRUS; HEPATITIS-C VIRUS; TRANSFERASE-ACTIVITY; STRAND RNA; SEQUENCE; INSECT; IDENTIFICATION; INITIATION; ORGANIZATION; PROTEINS;

D O I：

10.5483/BMBRep.2010.43.4.284

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Replication of positive-strand RNA virus is mediated by a virus-encoded RNA-dependent RNA polymerase (RdRp). To study the replication of Ectropis obliqua virus (EoV), a newly identified insect virus belonging to the family Iflaviradae, we expressed the RNA polymerase domain in Escherichia coli and purified it on a Ni-chelating HisTrap affinity column. It is demonstrated that EoV RdRp initiated RNA synthesis in a primer- and poly (A)-dependent manner in vitro. Furthermore, the effect of primer concentration, temperature, metal ions (Mg(2+), Mn(2+), and K(+)) on enzymatic activity were determined. Our study represented a first step towards understanding the mechanism of EoV replication. [BMB reports 2010; 43(4): 284-290]

引用

页码：284 / 290

页数：7

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