Immunodetection of the replicative complex of <Emphasis Type="Italic">Barley yellow dwarf</Emphasis><Emphasis Type="Italic">virus</Emphasis>-PAV <Emphasis Type="Italic">in vivo</Emphasis> |
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Authors: | VW?Fomitcheva Email author" target="_blank">J?SchubertEmail author F?Rabenstein A?Habeku? |
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Institution: | (1) Institute of Resistance Research and Pathogen Diagnostics, Germany;(2) Institute of Epidemiology and Resistance, Federal Centre for Breeding Research on Cultivated Plants,, Theodor-Roemer Weg 4, 06449, Aschersleben, Germany |
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Abstract: | The genomic fragments of two open reading frames (ORFs) 1 and 2 of German and Canadian PAV isolates of Barley yellow dwarf virus (BYDV-PAV) were sequenced. Sequences only slightly differed from previously published sequences of this virus. Two polyclonal antisera against proteins encoded by ORFs 1 and 2 of a German ASL-1 isolate were developed using recombinant antigens expressed in E. coli as a fusion either to His6− or thioredoxin-tags. In Western blot analysis with total protein extracts from BYDV infected plants, antisera efficiently recognized the 99 kDa fusion protein expressed from ORF1 and ORF2 (P1–P2 protein). Later in infection the P1–P2 protein disappeared and two smaller proteins, revealing sizes of 39 and 60 kDa, could be detected. |
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Keywords: | Barley yellow dwarf virus RNA-dependent RNA polymerase replication serology |
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