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首页> 外文期刊>Advancements in Life Sciences >Molecular detection, phylogenetic analysis and designing of siRNA against Potato Virus X
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Molecular detection, phylogenetic analysis and designing of siRNA against Potato Virus X

机译：马铃薯X病毒siRNA的分子检测，系统发育分析和设计

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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.
机译：背景：由于马铃薯（Solanum tuberosum L.）是最受人类饮食欢迎的粮食作物之一，因此增加其产量是科学家要实现的重要目标。在这项分子研究中，我们表征了马铃薯X病毒（PVX）的外壳蛋白（CP）基因。 CP基因是毒力的介体，是病毒结构装配的组成部分。方法：我们组织培养了PVX阳性马铃薯植株，用于病毒RNA的提取。将总RNA转化为cDNA，以在PCR中引发CP基因进行扩增。为了获得完整的基因序列，我们将CP基因克隆到pTZ57R / T克隆载体中。用EcoRI和HindIII限制酶双重消化重组质粒后，获得710bp的片段，证实了克隆。结果：推导的710 bp的共有序列为CP基因的准确cds，与参考基因组相似性为95％。系统发育分析表明，印度分离的PVX是最接近的一种。针对上述提到的问题设计了多个siRNA，并对其进行了计算优化，为进一步的研究提供了基础。 i）巴基斯坦PVX分离株的CP基因与世界各地发现的其他PVX分离株具有高度同源性; ii）确定有效si​​RNA介导的使PVX基因沉默的有效方法的靶点，这种保守的性质可以证明是非常有前景的。因此，通过siRNA介导的策略在巴基斯坦发展抗PVX的马铃薯作物，CP基因可能是最佳靶点。

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