Overexpression of Bombyx mori prothoracicotropic hormone using baculovirus vectors.
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.
O'Reilly
D R
DR
Department of Entomology, University of Georgia, Athens 30602, USA.
Kelly
T J
TJ
Masler
E P
EP
Thyagaraja
B S
BS
Robson
R M
RM
Shaw
T C
TC
Miller
L K
LK
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary; D026461:Ecdysteroids; D005786:Gene Expression Regulation; D017344:Genes, Insect; D005822:Genetic Vectors; D007301:Insect Hormones / Q000235:genetics* / Q000378:metabolism; D007814:Larva / Q000254:growth & development / Q000821:virology; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000235:genetics* / Q000378:metabolism; D017924:Nucleopolyhedroviruses / Q000235:genetics* / Q000472:pathogenicity; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D018411:Spodoptera / Q000196:embryology / Q000821:virology; D013256:Steroids / Q000378:metabolism
D018076:DNA, Complementary; D026461:Ecdysteroids; D007301:Insect Hormones; D009479:Neuropeptides; D011994:Recombinant Proteins; D013256:Steroids; C009089:prothoracicotropic hormone