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VP1, the putative virion-associated RNA-dependent RNA polymerase (RdRp) of infectious pancreatic necrosis virus (IPNV) can be guanylylated in vitro whereupon it becomes a primer for in vitro RNA synthesis [Virology 208 (1995) 19]. The role of a template or other virion polypeptides in the reaction is unknown. To shed light on this question, his-tagged recombinant VP 1 (rVP1) was expressed...
We have cloned and characterized the Drosophila X virus (DXV) genome segment B and its encoded VP1, the putative RNA-dependent RNA polymerase (RdRp) present in the virion. The 2991-bp open reading frame encodes the largest birnavirus VP1 at 977 aa, with a calculated M r of 112.8 kDa. As with the VP1 proteins of the type species of the other two genera in the family Birnaviridae, namely, infectious...
The RNA polymerase VP1 of IPNV (a bisegmented dsRNA containing virus) is present in the virion both as a free polypeptide and as a genome-linked protein (VPg). Virion VP1 primes viral RNA synthesisin vitro(P. Dobos, 1995,Virology208, 19–25), and here we present data which suggest that protein-primed RNA synthesis may also take place in infectious pancreatic necrosis virus (IPNV)-infected cells. Anti-VP1...
The dsRNA containing birnavirus, infectious pancreatic necrosis virus, possesses a virion-associated RNA-dependent RNA polymerase which acts both as primer and as polymerase duringin vitroRNA synthesis (P. Dobos, 1995,Virology208, 19–25). Using [α32P]GTP, we have radiolabeled virion RNAin vitroand found that after deproteinization most of the labeled product comigrated in agarose gels with the 3-kbp...
Drosophila X virus represents the entomobirnavirus genus of the Family Birnaviridae. Segment A of this bisegmented dsRNA containing virus was cloned and sequenced. The 3360-bp-long nucleotide sequence revealed the presence of two open reading frames (ORFs). A large ORF of 3096 nucleotides, which is flanked by a 107-bp 5′ and a 157-bp 3′-untranslated region, and a 711-nucleotide-long small ORF located...
A serotype-specific, conformational epitope of VP2 of infectious pancreatic necrosis virus (IPNV) (Jasper), has been mapped by restriction enzyme site-specific deletions of cloned viral cDNA. Subclones encoding fragments of VP2 were expressed in Escherichia coli followed by polyacrylamide gel electrophoresis and Western blot analysis. The epitope (between amino acid residues 242 and 325) reacted with...
A recombinant plasmid containing the Choristoneura fumiferana multinucleocapsid nuclear polyhedrosis virus (CfMNPV) HindIII R fragment (m.u. 2.2-3.9) was shown to undergo CfMNPV infection-dependent DNA replication in Cf-124T cells. Replication of this DNA sequence was detectable by 24 hr p.i. and did not appear to have resulted as a consequence of recombination with the virus genome. Replication was...
The 94-kDa virion-associated RNA-dependent RNA polymerase (RdRp) is present in infectious pancreatic necrosis virus in two forms: (i) as a free polypeptide (VP1) and (ii) as a genome linked protein (VPg) (J. G. Calvert et al., 1991, J. Gen. Virol. 72, 2563-2567). VP1 was guanylylated in vitro by incubating purified virus in the presence of [α 32 P]GTP. During further incubation in an in vitro...
IPNV is a medium-sized, unenveloped bisegmented dsRNA-containing virus in the family Birnaviridae. Genome segment A (3097 bp) contains two overlapping open-reading frames (ORFs). A large ORF encodes a 106 kDa polyprotein (NH2-pVP2-NS protease-VP3-COOH) which is cotranslationally cleaved by the protease to generate the major capsid proteins VP2 and VP3, and a second, small ORF which overlaps the...
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