PARTIAL MOLECULAR CHARACTERIZATION OF A CHINESE ISOLATE OF GRAPEVINE LEAFROLL-ASSOCIATED VIRUS 2 AND PRODUCTION OF ANTISERA TO RECOMBINANT VIRAL PROTEINS

Z.Y. Xu, N. Hong, B. Xing, G.P. Wang

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.

Keywords


Grapevine; Grapevine leafroll-asociated virus 2; gene cloning; sequencing; recombinant proteins; PAS-ELISA; Western blotting

Full Text:

PDF


DOI: http://dx.doi.org/10.4454/jpp.v88i1.835

Refbacks

  • There are currently no refbacks.


EDIZIONI ETS, Pisa, Italy