P34.8 (GP37) is not essential for baculovirus replication.
Previous reports have indicated that p34.8 (gp37) may be essential for the replication of Autographa californica nucleopolyhedrovirus (AcMNPV) because no virus with inactivated p34.8 was isolated. We have ascertained the requirement for this gene by attempting to inactivate it with a large insertion [the gene encoding GFP (green fluorescent protein)] or by deleting all the conserved domains from the open reading frame (ORF). The gene encoding GFP was inserted into the NOT:I site of the p34.8 ORF and a viral plaque containing the insertion was propagated in SF-21 cells. Similarly, 531 bp (NOT:I-XBA:I) containing all conserved domains were deleted from the ORF. All mutants were authenticated by PCR amplification, restriction endonuclease analysis, DNA sequencing, and Southern and Northern blot analysis. It was found that inactivation of p34.8 of AcUW1-LacZ (AcMNPV containing a lacZ gene in the p10 locus) had no effect on the biological property of virus, such as virulence and kinetics. These two independent methods showed that p34.8 is not essential for replication and that this locus could provide another site for the engineering of baculoviruses.
Cheng
Xiao-Wen
XW
Laboratory for Molecular Virology, Great Lakes Forestry Center, 1219 Queen St E, Sault Ste Marie, Ontario, CanadaP6A 5M71.
Krell
Peter J
PJ
Department of Microbiology, University of Guelph, Guelph, Ontario, CanadaN1G 2W12.
Arif
Basil M
BM
Laboratory for Molecular Virology, Great Lakes Forestry Center, 1219 Queen St E, Sault Ste Marie, Ontario, CanadaP6A 5M71.
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000737:chemistry / Q000235:genetics* / Q000502:physiology*; D001681:Biological Assay; D003465:Culex / Q000502:physiology / Q000821:virology; D020043:Genes, Essential / Q000235:genetics; D005818:Genetic Engineering; D007700:Kinetics; D007928:Lethal Dose 50; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D016366:Open Reading Frames / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
D012333:RNA, Messenger; D012367:RNA, Viral; D014764:Viral Proteins; C421744:p34.8 protein, Autographa californica nucleopolyhedrovirus