Insecticidal properties of genetically engineered baculoviruses expressing an insect juvenile hormone esterase gene

• Published in: Applied and Environmental Microbiology Vol. 58; no. 5; pp. 1583 - 1591
• Main Authors: Eldridge, R. (University of Georgia, Athens, GA), O'Reilly, D.R, Hammock, B.D, Miller, L.K
• Format: Journal Article
• Language: English
• Published: United States American Society for Microbiology 01.05.1992
• Subjects: Animals; AUTOGRAPHA CALIFORNICA; BACULOVIRIDAE; Baculoviridae - enzymology; Baculoviridae - genetics; Blotting, Western; Carboxylic Ester Hydrolases - biosynthesis; Carboxylic Ester Hydrolases - genetics; CONTROL BIOLOGICO; DESARROLLO BIOLOGICO; DEVELOPPEMENT BIOLOGIQUE; Enzyme Activation; ESTERASAS; ESTERASE; EXPRESION GENICA; EXPRESSION DES GENES; GAIN DE POIDS; GANANCIA DE PESO; GENE; Gene Expression; GENES; Genetic Engineering; Genetic Vectors; Genetics; HELIOTHIS VIRESCENS; HORMONAS JUVENILES; HORMONE JUVENILE; Hormones; Insects; Juvenile Hormones - genetics; Larva - microbiology; LUTTE BIOLOGIQUE; Moths - enzymology; Moths - genetics; Moths - microbiology; nuclear polyhedrosis virus; Pest Control, Biological - methods; RECOMBINACION; RECOMBINAISON; Viruses
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/aem.58.5.1583-1591.1992

Exploring the possibility of enhancing the properties of baculoviruses as biological control agents of insect pests, we tested the effect of expressing an insect gene (jhe) encoding juvenile hormone esterase. Juvenile hormone esterase inactivates juvenile hormone, which regulates the outcome of an insect molt. A cDNA encoding the juvenile hormone esterase of Heliothis virescens was inserted into the genome of Autographa californica nuclear polyhedrosis virus such that the gene was expressed under the control of a strong, modified viral promoter. This virus, however, naturally encodes an ecdysteroid UDP-glucosyltransferase which inactivates ecdysone, the hormone which initiates molting. Since ecdysteroid Udp-glucosyltransferase could mask the effects of jhe expression by blocking molting entirely, jhe-expressing viruses in which the ecdysteroid UDP-glucosyltransferase gene was deleted or disrupted were constructed. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of proteins from infected cells revealed several intracellular proteins and two major secreted proteins which reacted with antibodies to authentic juvenile hormone esterase. Western blot analysis coupled with tunicamycin treatment indicated that differential glycosylation was responsible for the multiple products. Hemolymph of recombinant virus-infected fourth-instar Trichoplusia ni larvae contained levels of juvenile hormone esterase activity 40-fold higher than maximal levels found in uninfected larvae. However, little or no difference in developmental characteristics, weight gain, or time of mortality was observed between insects infected with the jhe-expressing viruses and control viruses