Cloning of the Open Reading Frames of Potato Virus M, known as suppressors of RNAi.
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Keywords:
Potato virus M, RNA interference, RNA silencing suppressor proteins,Abstract
Gene silencing is an important regulatory and defense mechanism in plants. Plant viruses evolved some proteins that suppress cellular RNA interference mechanism. In this work identification of silencing suppressors of potato virus M (PVM) is reported. The cDNA sequences of different PVM open reading frames were cloned into the Gatewaycompatible binary T-DNA destination vector pMDC32, harboring a double 35S promoter. Each of these constructs was electroporated into Agrobacterium tumefaciens which then were agroinfiltrated into Nicotiana benthamiana 16C line. It was shown that viral coat protein (34K) as well as 12K protein possessed suppressor properties of cellular silencing activity. Thereafter, these open reading frames were cloned in both sense and antisense sequences into pUC19.References
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2 Voinnet O. Post-transcriptional RNA silencing in plant-microbe interactions: a touch of robustness and versatility //Curr.Opin. Plant Biol.- 2008.- V.11. - №4. - P.464-470.
3 Ruiz-Ferrer V., Voinnet O. Roles of plant small RNAs in biotic stress responses // Annual Review of Plant Biology.
– 2009. – V. 60. – P. 485-510.
4 Alvarado V., Scholthof H.B. Plant responses against invasive nucleic acids: RNA silencing and its suppression by
plant viral pathogens // Semin.Cell. Dev. Biol. – 2009. – V.20. - №9. – P.1032-1040.
5 Омаров Р.Т., Берсимбай Р.И. Биохимические механизмы супрессии РНК-интерференции вирусами растений // Биохимия. – 2010. – Т. 75. – Вып. 8. – С. 1062-1069.
6 Kryldakov R., Akbergenov R., Hohn T., Iskakov B. Identification of silencing suppressors of potato virus M // Journal of Cell and Molecular Biology . – 2011. – V.9. - №1. – P.15-20.
7 Zavriev S.K., Kanyuka K.V., Levay K.E. The genome organization of potato virus M RNA // J. Gen. Virol. – 1991.
– V. 72. – P. 9–14.
8 Proll E., Leiser R.M., Ostermann W.D., Spaar D. Some physicochemical properties of potato virus M // Potato Res.
– 1981. – V. 24. – P. 1–10.
9 Flatken S., Ungewickell V., Menzel W., Maiss E. Construction of an infectious full-length cDNA clone of potato virus M // Arch. Virol. – 2008. – V. 153. – P. 1385–1389.
10 Tavantzis S.M. Physicochemical properties of potato virus M // Virology. – 1984. – V. 133. – P. 427-430.
11 Sozinova L., Abay A., Ramankulov E., Nachmias A., Loebenstein G. Reducing Virus Incidence in Potato Seed Tubers by Rapid Propagation: A Pilot Scheme in Kazakhstan // Phytoparasitica. – 2007. – V.35. - № 2. – P.201.
12 Curtis M.D., Grossniklaus U. A Gateway Cloning Vector Set for High-Throughput Functional Analysis of Genes in
Planta // Plant Physiology. – 2003. – V.133. – P.462-469.
13 Voinnet O., Pinto Y.M., Baulcombe D.C. Suppression of gene silencing: A general strategy used by diverse DNA
and RNA viruses of plants // PNAS. – 1999. – V.96. - № 24. – P. 14147-14152.
14 Maniatis T., Fritsch E.F., Sambrook J. Molecular cloning. A laboratory manual - 1ed.-New York: Cold Spring Harbor
Lab. Press, 1989. - 3 vols.
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Kryldakov, R., Iskakov, B., Stanbekova, G., & Karpova, O. (2015). Cloning of the Open Reading Frames of Potato Virus M, known as suppressors of RNAi. Experimental Biology, 60(2), 274–277. Retrieved from https://bb.kaznu.kz/index.php/biology/article/view/952
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