Journal of Animal and Veterinary Advances

Year: 2012
Volume: 11
Issue: 13
Page No. 2316 - 2322

Construction and Characterization of an Expression Plasmid Containing Peste des Petits Ruminants Virus Phosphoprotein Gene

Authors : J.J. Zhai, Y.X. Dou, X.N. Luo and X.P. Cai

Abstract: The Phosphoprotein (P) of genus Morbillivirus in the family Paramyxoviridae is an essential component of the viral RNA Polymerase complex (P-L) and is thought to mediate the interaction of the large protein (L) with the viral nucleocapsid template for transcription and replication. To date, the precise role of Peste des Petits Ruminants Virus (PPRV) P is unclear. To establish its function in the present study, the encoding sequence of PPRV P was obtained by RT-PCR and then inserted into the multiple cloning site of pcDNA3.1 vector. The constructed recombinant plasmid was successfully transfected into the Vero cell as confirmed by indirect immunofluorescence assay, SDS-PAGE and Western blotting. Then, the transfected cells were inoculated by PPRV. The results showed that the constructed plasmid which was designated pcDNA 3.1/P could be expressed in Vero cells. After inoculation with PPRV, Cytopathic Effects (CPE) have been detected in the transfected cells by plasmid pcDNA3.1/P a day earlier than in cells of the control group. In conclusion, the results showed that the phosphoprotein did accelerate the virus replication; this would be very useful for further study of P protein functions during PPRV transcription and replication.

How to cite this article:

J.J. Zhai, Y.X. Dou, X.N. Luo and X.P. Cai, 2012. Construction and Characterization of an Expression Plasmid Containing Peste des Petits Ruminants Virus Phosphoprotein Gene. Journal of Animal and Veterinary Advances, 11: 2316-2322.

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