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International Journal of Virology

Year: 2005  |  Volume: 1  |  Issue: 1  |  Page No.: 3 - 3

Cloning, exprision and nucleotide sequence of coat protein gene of an egyptian isolate of potato virus Y strain NTN infecting potato plants

M.A.Amer, M. H. El-Hammady, H.M. Mazyad, A. A.Shalaby and F.M. Abo-El- Abbas

Abstract

The coding sequence of coat protein gene of an Egyptian isolate of PVY was modified to has an initation AUG code for translation. Reverse Transcription-Polymerase Chain Reaction (RT-PCR) amplified product of an expected size of 801 bp, cloned into the Pin Point Xa-1 protein expression vector. The fidelity of each PCR amplified PVY-CP gene was tested by PCR, restriction analysis and translation. Analysis by in vitro translation using western blotting assay on nitrocellulose membrane using monoclonal antibodies verified that the PVY-CP gene correctly encoded and expressed a protein reacting with PVY antibodies. The CP gene ( 32 KDa) was reacted successfully with PVY specific monoclonal antibodies in dot blot immunobinding assay ( DIBA) as well as in western blot assay. The nucleotide sequence of the Egyptian isolate of PVY CP gene was determined to be 801 nucleotides in length encoding a deduced amino acid. Nuelcotide sequence analysis revealed a range of 96- 99.5% sequence identity among the PVY NTNisolates.

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