Cloning, Expression and Nucleotide Sequence of Coat Protein Gene of an Egyptian Isolate of Potato Virus y Strain NTN Infecting Potato Plants
Cloning, Expression and Nucleotide Sequence of Coat Protein Gene of an Egyptian Isolate of Potato Virus y Strain NTN Infecting Potato Plants
M. A. Amer1; M. H. El-Hammady2; H. M. Mazyad1; A. A. Shalabyl and F. M. Abo-El-abbas
ABSTRACT
Reverse transcription- polymerase chain reaction (RT-PCR) amplified product of the expected size of 801 bp was cloned into the Pinpoint Xa-l protein expression vector. The accury 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, approximately 32 KDa reacted successfully with P VY 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 267 amino acid. Nucleotide sequence analysis revealed a range of 96- 99.5 % sequence identity among the PVYNTN isolate (PVY-T, PVY SWS, PVY II and PVY NTN).
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