Expression of polydnavirus genes under polydnavirus promoter regulation in insect larvae infected with baculovirus recombinants.
We have evaluated the use of baculoviruses to deliver Campoletis sonorensis polydnavirus (CsPDV) genomic DNA into lepidopteran larvae to facilitate the identification of functional CsPDV genes. Genomic fragments consisting of regulatory (promoter) and coding sequences for two CsPDV genes (VHv1.1 and WHv1.6) were used to generate CsPDV-baculovirus recombinants and evaluate the expression of genes under the regulation of the CsPDV promoters. Northern blot and primer extension studies established that CsPDV genes were expressed under the control of their own promoters in these CsPDV-baculovirus recombinants. Transcripts were detected as early as 4 h post-infection indicating that temporal activity of CsPDV promoters was retained. The VHv1.1 gene product as expressed from CsPDV-baculovirus recombinants was identical in size and in functional properties to that produced in CsPDV-infected insects. CsPDV-baculovirus recombinants may be useful for the screening and characterization of polydnavirus genes with functional activities that can only be evaluated in insect larvae.
Soldevila
A I
AI
Department of Entomology, University of Kentucky, Lexington, KY 40546-0091, USA.
Webb
B A
BA
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004279:DNA, Viral; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D006434:Hemocytes / Q000378:metabolism; D007814:Larva; D008969:Molecular Sequence Data; D009036:Moths / Q000166:cytology / Q000821:virology; D017924:Nucleopolyhedroviruses / Q000235:genetics; D018071:Polydnaviridae / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins; D018411:Spodoptera / Q000166:cytology; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism
D004279:DNA, Viral; D011993:Recombinant Fusion Proteins; C099400:VHv1.1 protein, Campoletis sonorensis polydnavirus; D014764:Viral Proteins