biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 46:253-260, 2003 | DOI: 10.1023/B:BIOP.0000022260.95561.1d

Sequence Variability of Helper Component Protein of Potato Virus Y Identified by Thermodynamic Methods

J. Škopek1,2, J. Matoušek1
1 Institute of Plant Molecular Biology, Academy of Sciences of the Czech Republic, České Budějovice, Czech Republic
2 Faculty of Biological Sciences, University of South Bohemia, České Budějovice, Czech Republic

An extent of helper component protein (HC-Pro) sequence variability within virus population of single Czech isolate of potato virus YNTN (PVYNTN) Nicola was identified by temperature-gradient gel electrophoresis (TGGE) and heteroduplex analysis. HC-Pro region was approximated with 6 pairs of primers derived from Hungarian PVYNTN isolate (sequence AC M95491). Immunocapture reverse transcription - polymerase chain reaction (RT-PCR) was used to obtain six mixtures of individual overlapping polymerase chain reaction (PCR) products. cDNA libraries were prepared by cloning of purified PCR products in pCR-Script vector and screened by heteroduplex analyses. 15 different subfragments within the HC-Pro region were isolated and sequenced. In comparison to AC M95491, 19 nucleotide changes were identified, 13 led to amino acid (aa) changes. In comparison to available post-transcriptional gene silencing (PTGS) specific suppressor HC-Pro sequences of potyviruses we found out 4 aa changes in conserved regions.

Keywords: cDNA analysis; heteroduplex analysis; PTGS suppressor; TGGE
Subjects: amino acids; cDNA analysis; helper component protein, potato virus Y; heteroduplex analysis; polymerase chain reaction (PCR); post-transcriptional gene silencing suppressor; potato virus Y; potyvirus; reverse transcriptase polymerase chain reaction (RT-PCR); temperature-gradient gel electrophoresis; tobacco etch virus

Published: September 1, 2003  Show citation

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Škopek, J., & Matoušek, J. (2003). Sequence Variability of Helper Component Protein of Potato Virus Y Identified by Thermodynamic Methods. Biologia plantarum, 46(2), 253-260. doi: 10.1023/B:BIOP.0000022260.95561.1d
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