NDLI: The VP2 protein of grass carp reovirus (GCRV) expressed in a baculovirus exhibits RNA polymerase activity

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Virologica Sinica : Volume 29, Issue 1, February 2014

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The VP2 protein of grass carp reovirus (GCRV) expressed in a baculovirus exhibits RNA polymerase activity

Content Provider	Springer Nature Link
Author	Yan, Liming Liu, Huan Li, Xiaoming Fang, Qin
Copyright Year	2014
Abstract	The double-shelled grass carp reovirus (GCRV) is capable of endogenous RNA transcription and processing. Genome sequence analysis has revealed that the protein VP2, encoded by gene segment 2 (S2), is the putative RNA-dependent RNA polymerase (RdRp). In previous work, we have ex-pressed the functional region of VP2 that is associated with RNA polymerase activity (denoted as rVP2$^{390–900}$) in E. coli and have prepared a polyclonal antibody against VP2. To characterize the GCRV RNA polymerase, a recombinant full-length VP2 (rVP2) was first constructed and expressed in a baculovirus system, as a fusion protein with an attached His-tag. Immunofluorescence (IF) assays, together with immunoblot (IB) analyses from both expressed cell extracts and purified Histagged rVP2, showed that rVP2 was successfully expressed in Sf9 cells. Further characterization of the replicase activity showed that purified rVP2 and GCRV particles exhibited poly(C)-dependent poly(G) polymerase activity. The RNA enzymatic activity required the divalent cation Mg$^{2+}$, and was optimal at 28 °C. The results provide a foundation for further studies on the RNA polymerases of aquareoviruses during viral transcription and replication.
Starting Page	86
Ending Page	93
Page Count	8
File Format	PDF
ISSN	16740769
Journal	Virologica Sinica
Volume Number	29
Issue Number	2
e-ISSN	1995820X
Language	English
Publisher	Wuhan Institute of Virology, CAS
Publisher Date	2014-03-04
Publisher Place	Heidelberg
Access Restriction	Subscribed
Subject Keyword	grass carp reovirus (GCRV) VP2 protein baculovirus recombinant RNA polymerase Virology Medical Microbiology Oncology Biochemistry Microbiology Microbial Genetics and Genomics
Content Type	Text
Resource Type	Article
Subject	Infectious Diseases Virology Immunology Molecular Medicine

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