אסיף מאגר המחקר החקלאי

The use of S1 nuclease treatment of hybrid PCR products for the differentiation between PVY isolates

Year:

1998

Source of publication :

Authors :

Volume :

132

Co-Authors:

Rosner, A., Department of Virology, Agricultural Research Organization, Volcani Center, Bet-Dagan 50250, Israel
Spiegel, S., Department of Virology, Agricultural Research Organization, Volcani Center, Bet-Dagan 50250, Israel
Maslenin, L., Department of Virology, Agricultural Research Organization, Volcani Center, Bet-Dagan 50250, Israel
Levy, D., Department of Field Crops, Agricultural Research Organization, Volcani Center, Bet-Dagan 50250, Israel

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Abstract:

A new strategy based on treating PCR hybrids with S1 nuclease was used to differentiate between two PVY isolates. Mixed denatured and annealed hybrid PCR products of two PVY isolates including a tested strain and a reference N strain were treated with S1 nuclease. Single-stranded mismatched regions were revealed by the S 1 nuclease cleavage, yielding a characteristic pattern of bands in polyacrylamide gel by which virus isolates could be matched. Sequence analysis of the relevant PCR products revealed that only part of the mismatched regions were cleaved by the S 1 nuclease. Still, the distinct pattern of degradation products enabled the differentiation between the PVY isolates. The general application of this procedure for strain differentiation is discussed.

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