Differentiating PVY(NTN) from PVY(N) by annealing to reference RNA transcripts.
A method for the differentiation of virus strains based on the shift in electrophoretic mobility of partially annealed RNA transcripts is described. Oppositely oriented RNA transcripts of the NTN- and N-strains of PVY, complementary at their 3'-end variable (strain-specific) region, were annealed to form a partial duplex which moved more slowly in gel than heterologous (NTN+N) unpaired transcripts. Thus, the two virus strains could be identified by annealing to a known reference transcript. The rate of duplex migration was correlated with transcript lengths and could be tightly controlled thereby. Thus, a higher degree of resolution was obtained than with transcript conformation polymorphism, which is empirical and unpredictable in nature.
Rosner
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A
Department of Virology. The Volcani Center, 50250, Bet Dagan, Israel. rosnera@agri.gov.il
Maslenin
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L
D014644:Genetic Variation; D016679:Genome, Viral; D009693:Nucleic Acid Hybridization; D018515:Plant Leaves / Q000821:virology; D016133:Polymerase Chain Reaction; D017800:Potyvirus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics*; D012015:Reference Standards; D012680:Sensitivity and Specificity; D011198:Solanum tuberosum / Q000821:virology; D014158:Transcription, Genetic
D012333:RNA, Messenger; D012367:RNA, Viral