A simple and efficient procedure for the oral inoculation of Trichoplusia ni larvae with polyhedrin-negative recombinant baculovirus.
Current procedures for inoculating lepidopteran larvae with polyhedrin-negative recombinant baculovirus, i.e. intracoelomic injection or coinfection with wild type virus, are laborious and can compromise final yields of recombinant protein. Herein is described a simple and efficient method for oral inoculation. Up to 100% infection was obtained when individual early fifth instar Trichoplusia ni larvae were fed a small piece of a formaldehyde-free insect diet to which 4.2 x 10(5) PFU of a polyhedrin-negative recombinant Autographa californica nuclear polyhedrosis virus (AcNPV) containing the gene for beta-galactosidase was applied. Infected larvae were identified by assaying hemolymph for beta-galactosidase activity. The maximum levels of beta-galactosidase detected in these hemolymph samples were identical to those obtained for larvae infected by intracoelomic injection. The dose of polyhedrin-negative recombinant virus recommended for intracoelomic injection of T. ni was efficacious for the oral route of inoculation.
Department of Plant Pathology, University of Kentucky, Lexington.
J F
JF
van den Heuvel
D W
DW
Thornbury
T P
TP
Pirone
J A
JA
Lesnaw
D000284:Administration, Oral; D000818:Animals; D016367:Baculoviridae / Q000254:growth & development*; D004274:DNA, Recombinant / Q000235:genetics; D007814:Larva / Q000382:microbiology; D007915:Lepidoptera / Q000382:microbiology*; D008828:Microbiological Techniques; D000077183:Occlusion Body Matrix Proteins; D011994:Recombinant Proteins / Q000096:biosynthesis; D013217:Starvation; D014764:Viral Proteins / Q000235:genetics; D015678:Viral Structural Proteins; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D004274:DNA, Recombinant; D000077183:Occlusion Body Matrix Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C009829:polyhedrin protein, Nucleopolyhedrovirus; D001616:beta-Galactosidase