Abstract
Background: As potato (Solanum tuberosum L.) is one of the most liked food crops for
human diet so increasing its production is an important goal for scientists to achieve. In this
molecular study, we characterized the Coat Protein (CP) gene of Potato Virus X (PVX). CP
gene is virulence mediator and integral part of viral structural assembly.
Methodology: We tissue cultured the PVX positive potato plants for viral RNA extraction.
Total RNA was converted to cDNA for priming CP gene in PCR for amplification. To get the
complete sequence of gene, we cloned CP gene into pTZ57R/T cloning vector. Upon double
digestion of recombinant plasmid with EcoRI and HindIII restriction enzymes, 710 bp
fragment was obtained which confirmed cloning. Recombinant plasmid was sequenced with
M13 primers.
Results: Derived consensus sequence of 710 bp was found to be exact cds of CP gene
showing 95% similarity with referenced genome. Phylogenetic analysis suggested Indian
isolate of PVX as the nearest one. Multiple siRNA were designed against mentioned and
optimized computationally to provide base for further studies.
Conclusion: Following facts may be established upon findings of this research; i) CP gene
of Pakistani isolate of PVX has high homology with other PVX isolates found around the
world, ii) in determining target for efficient siRNA mediated approach to silence PVX
genome, this conserved nature can be proved very promising. Thus, to develop PVX-resistant
potato crop in Pakistan through siRNA mediated strategy, CP gene could be the best target.
Shafique Ahmed, Idrees Ahmed Nasir, Tahir Yaqub, Muhammad Waseem, Bushra Tabassum, Faiza Masood, Anwar Khan, Shahid Javed Butt, Tayyab Husnain. (2013) Molecular detection, phylogenetic analysis and designing of siRNA against Potato Virus X, Advancements in Life Sciences, Volume 1 , Issue 1.
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