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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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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)
  • Published : 2010.04.30
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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.

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References

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