Overexpression of Bombyx mori prothoracicotropic hormone using baculovirus vectors

• Published in: Insect biochemistry and molecular biology 1995-04, Vol.25 (4), p.475-485
• Main Authors: O'reilly, David R., Kelly, Thomas J., Masler, Edward P., Thyagaraja, Belgaum S., Moy Robson, R., Shaw, Terence C., Miller, Lois K.
• Format: Article
• Language: English
• Subjects: AcMNPV; Amino Acid Sequence; Animals; Autographa californica nuclear polyhedrosis virus; baculovirus; Base Sequence; Bombycidae; Bombyx - genetics; Bombyx mori; Bombyx mori Spodoptera frugiperda; Cloning, Molecular; DNA, Complementary; Ecdysteroids; Gene Expression Regulation; Gene expression vectors; Genes, Insect; Genetic Vectors; Insect Hormones - genetics; Insect Hormones - metabolism; Insect-virus interactions; Larva - growth & development; Larva - virology; Lepidoptera; Molecular Sequence Data; Neuropeptides - genetics; Neuropeptides - metabolism; Nucleopolyhedrovirus - genetics; Nucleopolyhedrovirus - pathogenicity; Prothoracicotropic hormone; PTTH; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Spodoptera - embryology; Spodoptera - virology; Steroids - metabolism; Viral pathogenesis
• ISSN: 0965-1748; 1879-0240
• DOI: 10.1016/0965-1748(94)00087-F

Recombinant baculoviruses were constructed that express the cDNA encoding the prothoracicotropic hormone (PTTH) of Bombyx mori. This hormone stimulates the production of ecdysteroids by the insect's prothoracic glands. Two groups of viruses were constructed, expressing either the entire cDNA encoding prepro-PTTH, or a synthetic chimeric gene encoding a signal peptide fused to the mature PTTH subunit. In both cases, the genes were expressed in wild-type Autographa californica nuclear polyhedrosis virus (AcMNPV) and in vEGTDEL, an AcMNPV mutant that lacks a functional egt gene. The egt gene is required for viral-mediated inactivation of host ecdysteroids. High levels of functional PTTH were produced only by viruses expressing the mature subunit cDNA. This recombinant PTTH resembled the native hormone by all criteria examined. The overproduction of B. mori PTTH induced higher than normal levels of haemolymph ecdysteroids but had no observable effects on the development of infected Spodoptera frugiperda larvae. However, expression of PTTH by AcMNPV was found to inhibit the pathogenicity of the virus. This effect was particularly marked in the case of viruses lacking a functional egt gene.