We have sequenced the genome of a North American isolate of Kashmir bee virus (KBV). The sequence data show that KBV is closely related to acute bee paralysis virus (ABPV; GenBank accession AF150629) across the genome, with about 85% identity at amino acid level and about 80% identity at nucleotide level. It still remains to be determined if KBV is a separate virus species, or a strain of ABPV, based on these sequence data. The Kashmir bee virus genome has two major open reading frames, each encoding a polyprotein that presumably is subsequently processed into its active enzymatic constituents. In order to determine the pattern of genome expression, the polyprotein open reading frames were subcloned into a bacterial protein expression vector. The purpose of subcloning is to produce proteins that are specific to KBV. Then, we purify the proteins to make monoclonal antibodies, each of which will be specific for a single epitope of the polyprotein, and therefore useful for determining the pattern of polyprotein processing and the expression profile of each protein fragment. Antibodies against non-structural proteins (those involved in replication and polyprotein processing) will also be used to determine the sites of virus replication within honeybees and (possibly) varroa mites, through immuno-(electron)microscopy.
Species 1: Hymenoptera Apidae Apis mellifera (honey bee)
Keywords: cloning, protein
The ESA 2001 Annual Meeting - 2001: An Entomological Odyssey of ESA