Abstract
Solenopsis invicta virus 1 (SINV-1) is the first virus to be discovered and characterized in any ant species (Valles et al. 2004). The virus exhibits a host range limited to fire ants in the Solenopsis genus (Valles et al. 2007). The single-stranded RNA genome is composed of 8,026 nucleotides containing 2 large open reading frames (ORFs) in the sense orientation with an untranslated region (UTR) at each end and between the two ORFs. The 5’-proximal ORF encodes for the non-structural proteins, helicase, protease, and RNA-dependent RNA polymerase (RdRp) and the 3’-proximal ORF for the structural (or capsid) proteins. ORF 2 was originally reported to commence at nucleotide position 4,390 at a canonical AUG start codon. However, it was later revealed empirically to actually start at codon GCU (genome position 4423-4425) which encodes for an alanine (Valles and Hashimoto 2008). SDS-PAGE analysis of purified SINV-1 particles yields 3 major (VP1, VP2, VP3) and one minor (VP4) capsid proteins. N-terminal analysis has revealed the margins of the capsid proteins and the corresponding scissile bonds within the structural polyprotein (Valles and Hashimoto 2008). Amino acid residues at these junctions were consistent with other dicistroviruses and unclassified picorna-like insect-infecting viruses (Liljas et al. 2002). The objective of this short communication was to confirm production and genome location of VP3 within the SINV-1 structural polyprotein using Western analysis. Polyclonal antibodies were synthesized from a recombinant peptide sequence obtained from the predicted amino acid sequence of VP3 (i.e., SRGGYRYKFFADDN). The peptide sequence is located at amino acid positions 9971010 of the structural polyprotein (Fig. 1A). Polyclonal antibodies were raised in a rabbit host by GenScript USA, Inc. (Piscataway, NJ) according to the company’s standard protocols. Ants used for purification of virus were collected by excavating nests from locations in Gainesville, FL. Colonies were examined by RT-PCR methods to identify ants exclusively infected with
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