Nucleotide sequencing and transcriptional mapping of the Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus...

A 32P-labeled cloned DNA fragment (AcMNPV HindIII-Q) containing one of the repeated sequences and a portion of the p10 gene from Autographa californica multicapsid nuclear polyhedrosis virus (AcMNPV) was used to probe Southern blots containing restriction endonuclease digests of Orgyia pseudotsugata muticapsid nuclear polyhedrosis virus (OpMNPV) DNA. A single 3.6-kb fragment, OpMNPV HindIII-Q, was hybridized. The OpMNPV HindIII-Q fragment was cloned into pUC-18, mapped with restriction endonucleases, and reprobed with the AcMNPV HindIII-Q fragment. A small region of ca. 700 bp, near the left end of the cloned fragment, was cross-hybridized. DNA sequencing in this region revealed an open reading frame of 279 bp which shares detectable homology with the p10 gene of AcMNPV. The sequences downstream from the p10 gene in both viruses also contain long open reading frames which share homology. Northern blot analysis of RNA from OpMNPV infected O. leucostigma cells was used to define the temporal and spatial organization of transcripts from this region. S1 analysis of both termini of the major p10 mRNA indicates nontranslated regions of 52-53 bases at the 5' end and 175 bases at the 3' end. The 5'-mRNA start site was located within a 12-nucleotide sequence which is conserved in all late hyperexpressed baculovirus genes.