Pakistan Journal of Biological Sciences1028-88801812-5735Asian Network for Scientific Information10.3923/pjbs.2008.1482.1486Potato Y potyvirus in Potato Farms of Kermanshah using RT-PCR Amplification of the P1- Protease Gene and Its Cloning]]>MirmomeniM.H. SharifiA. SisakhtnezhadS. 1120081111In this study, the RT-PCR method was used to detect the Y virus
in potato tubers and leaves. Samples suspicious of virus infection with
symptoms of virus infection were gathered from farms in Kermanshah and
placed in plastic bags and kept at -80 °C temperature in order to maintain
the RNA of the virus until extraction. The extraction and purification
of RNA were carried out using Tri-Reagent kit. One of the virus genes
is the P1 protease gene which codes a proteinase enzyme. This enzyme plays
a role in breaking the initial polyprotein. For amplification of this
gene three primer, including primer-1, primer-2 and primer-3, were designed
and used. Using primer 1 and reverse transcriptase enzyme, cDNA was synthesized
and then PCR was performed using the primers 1, 2 and 3. The PCR products
were examined by agarose gel electrophoresis (1%). Consequently, two pieces
of DNA (400 and 800 bp) were yielded which were identical to the genome
DNA sequencing. Thus, the proposed technique is a convenient method for
quick and accurate detection of viruses and, therefore, the application
of this method for detecting Potato Y virus in potato farms is
recommended.]]>Akad, F. and H. Czosnek,2002Potato Virus Y (PVY) and PVYN, Method: RT-PCR.]]>2002Akad, F. and H. Czosnekh,2005Potato Virus X (PVX). Method: Immunocapture RT-PCR.]]>2005Boonham, N., K. Walsh, M. Hims, S. Preston, J. North and I. Barker,2002Potato virus Y isolates associated with potato tuber necrotic ring spot disease.]]>51117126Carrington, J.C., P.E. Jensen and M.C. Schaad,199814393400Ghosh, S.B., L.H.S. Nagi, T.R. Ganapathi, S.M.P. Khurana and V.A. Bapat,200282855859Lorenzen, J.H., T. Meacham, P.H. Berger, P.J. Shiel, M. Crosslin, P.B. Hamm and H. Kopp,2006Potato virus Y isolates collected in the Western USA and their comparison to isolates from Europe and Canada.]]>15110551074Maki-Valkama, T.K.,2000Potato virus Y (PVY) in potato cv. Pito transformed with the P1 gene of PVY.]]>2000Mestre, P., G. Brigneti, M.C. Durrant and D.C. Baulcombe,2003Potato virus Y NIa protease activity is not sufficient for elicitation of RY-mediated disease resistance in potato.]]>36755761Moran, J. and B. Rodoni,2003Potato virus Y.]]>Moravec, T., N. Cerovska and N. Boonham,2003NTN) using a three-primer PCR based in the coat protein gene.]]>1096368Moury, B., C. Morel, E. Johansen and M. Jacquemond,2002Potato virus Y and in the coat protein of other potyviruses.]]>8325632573Sambrook, J. and D.W. Russell,20013rd Edn.,Pages: 2344Pages: 2344Schoen, C.D. and G. Leone,19981998Shalaby, A. and H. Mazyad,20022002Spetz, C.,20032003Syller, J.,200667119130Urcuqui-Inchima, S., A.L. Haenni and F. Bernardi,200174157175Verchot, J. and J.C. Carrington,19956936683674Arbatova, J., K. Lehto, E. Pehu and T. Pehu,1998Potato Y Potyvirus in association with cytoplasmic inclusion bodies and in the cytoplasm of infected cells.]]>7923192323Varveri, C.,200028141148