Detection of baculovirus gene expression in insect cells and larvae by low light video image analysis.
Recombinant baculovirus isolates BmNPVluc and AcNPVluc (Kopylova-Sviridova et al., 1990) expressing the luc gene in Bombyx mori N-4, and in Sf 9 and Trichoplusia ni 368 cells, respectively, were studied. Luc gene expression driven by baculovirus regulatory elements was detected by enzyme and photometric assays. The expression of recombinant AcNPVluc and BmNPVluc genes in infected larvae of the cabbage looper, T. ni and the tomato hornworm Manduca sexta was analyzed by low-light video image methods. Expression of the luc gene was detected at high levels in both the lepidopteran cells and in third to fifth instar T. ni larvae. However, no light emission was detected in M. sexta caterpillars. High levels of light emission were detected in T. ni larvae when occlusion bodies containing both wild type and recombinant virus were fed to larvae. The results of these experiments demonstrate that video image analysis can be used to monitor the progression of baculovirus infection in susceptible insect cells and larvae. Bioluminescence in recombinant virus infected larvae can be used to determine virus host range, to monitor latent virus infection in insect cells and to assess the spread of recombinant viruses in the environment. Video image analysis was found to be a sensitive method for rapid detection and semiquantitative measurement of luc gene expression in baculovirus infected cells and for monitoring virus infection in larval tissues.
Langridge
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WH
Department of Microbiology and Molecular Genetics, Loma Linda University, CA 92350, USA.
Krausova
V I
VI
Szalay
A A
AA
Fodor
I
I
D000818:Animals; D012831:Bombyx / Q000166:cytology / Q000821:virology; D002460:Cell Line; D015870:Gene Expression; D005822:Genetic Vectors; D007089:Image Enhancement; D008156:Luciferases / Q000235:genetics*; D018402:Manduca / Q000378:metabolism; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D018411:Spodoptera / Q000166:cytology / Q000378:metabolism; D014741:Video Recording
D008156:Luciferases