Archivio Journal of Plant Pathology

Abstract:

The genes encoding RNA-dependent RNA polymerase (RdRp) and coat protein (CP) of a Chinese isolate of Grapevine leafroll-associated virus 2 (GLRaV-2- CF) were cloned in the vector pET-28a (+) and sequenced. Comparisons of these sequences with those of other GLRaV-2 isolates available from databases revealed homology levels with GLRaV-2-PN and GLRaV- 2-Sem higher than 98% at the nucleotide and amino acid levels. GLRaV-2-CF had lower homology with GLRaV-2-RG, another distinct strain, with similarities of 78% (RdRp) and 79% (CP) at the nucleotide level. The cloned RdRp and CP were successfully expressed in Escherichia coli BL-21 (DE3 plysS) and the resulting recombinant proteins were used to raise antisera in rabbits. Tests using these antisera readily detected the recombinant proteins in Western blots and the native virus proteins in infected grapevine samples.