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http://dx.doi.org/10.5483/BMBRep.2010.43.4.284

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

Lin, Meijuan (State Key Laboratory of Virology, College of Life Sciences, Wuhan University)
Ye, Shan (State Key Laboratory of Virology, College of Life Sciences, Wuhan University)
Xiong, Yi (State Key Laboratory of Virology, College of Life Sciences, Wuhan University)
Cai, Dawei (State Key Laboratory of Virology, College of Life Sciences, Wuhan University)
Zhang, Jiamin (State Key Laboratory of Virology, College of Life Sciences, Wuhan University)
Hu, Yuanyang (State Key Laboratory of Virology, College of Life Sciences, Wuhan University)
Publication Information
BMB Reports / v.43, no.4, 2010 , pp. 284-290 More about this Journal
Abstract
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.
Keywords
Ectropis obliqua virus; Iflavirus; Primer-dependent; RNA-dependent RNA polymerase; RNA synthesis;
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