ISHS Acta Horticulturae 657: XIX International Symposium on Virus and Virus-like Diseases of Temperate Fruit Crops - Fruit Tree Diseases
PRODUCTION AND USE OF POLYCLONAL ANTIBODIES TO THE COAT PROTEIN OF APPLE STEM GROOVING VIRUS EXPRESSED IN ESCHERICHIA COLI |
| Authors: | O. Nickel, T.V. Fajardo, A.P. Trivilin, M.L.N.P. Targon, M.A. Machado |
| Keywords: | Apple, ASGV, ELISA, antibodies, recombinant viral antigen |
| Abstract:
The coat protein (cp) gene of Apple stem grooving Capillovirus was amplified by RT-PCR, cloned, sequenced and subcloned in the expression plasmid pMal-c2. The ASGV cp gene was expressed in E. coli as a fusion protein (fp) containing a fragment of E. coli maltose binding protein (MBP). Bacterial cells were disrupted by sonication and the ASGVcp/MBP-fp was purified by amylose resin affinity chromatography. Polyclonal antibodies from rabbits immunized with the fp, gave specific reactions to ASGV from several infected apple cultivars at dilutions of up to 1:2000 in indirect ELISA. The ASGVcp/MBP-fp reacted to antisera raised against it as well as to commercial antisera against ASGV virions in immunoblotting. These results are a fundamental contribution to the development of an efficient ASGV-indexing of propagative and mother stock materials in Southern Brazil. | |
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