A. Rosner et al., THE USE OF S1 NUCLEASE TREATMENT OF HYBRID PCR PRODUCTS FOR THE DIFFERENTIATION BETWEEN PVY ISOLATES, Annals of Applied Biology, 132(1), 1998, pp. 107-114
A new strategy based on treating PCR hybrids with S1 nuclease was used
to differentiate between two PVY isolates. Mixed denatured and anneal
ed hybrid PCR products of two PVY isolates including a tested strain a
nd a reference N strain were treated with S1 nuclease. Single-stranded
mismatched regions were revealed by the S1 nuclease cleavage, yieldin
g a characteristic pattern of bands in polyacrylamide gel by which vir
us isolates could be matched. Sequence analysis of the relevant PCR pr
oducts revealed that only part of the mismatched regions were cleaved
by the S1 nuclease. Still, the distinct pattern of degradation product
s enabled the differentiation between the PVY isolates. The general ap
plication of this procedure for strain differentiation is discussed.