Expression of hornet genes encoding venom allergen antigen 5 in insects.
Antigen 5, also known as Dol m V, is a major allergen found in the venom of the baldfaced hornet, Dolichovespula maculata. We have inserted the f10 and f17 cDNAs, which encode hornet antigen 5 (HA5) forms 2 (Dol m; VB) and 3 (Dol m VA), respectively, into the genome of the baculovirus, AcMNPV, to produce the recombinant baculovirus gene expression vectors, vEV-HA5f10 and vEV-HA5f17. Insect cells infected with either vEV-HA5f10 or vEV-HA5f17 produce and secrete a novel protein with an electrophoretic mobility which is similar if not identical to authentic mature Dol m V. The gene products also react specifically with a polyclonal antiserum raised to Dol m VB as expected. Dol m V gene products were not acutely toxic when injected into insect larvae. However, infection of fifth instar larvae with vEV-HA5f17 resulted in premature melanization of the larvae and lower weight gain than infection with control virus. Thus, the Dol m V gene product has a subtle, possibly cytotoxic or biochemical effect on insects. The expression systems may prove useful in further structural and functional characterization of these proteins.
Tomalski
M D
MD
Department of Entomology, University of Georgia, Athens 30602.
King
T P
TP
Miller
L K
LK
D000485:Allergens / Q000235:genetics* / Q000633:toxicity; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D007313:Insecta / Q000187:drug effects; D007814:Larva / Q000187:drug effects; D008543:Melanins / Q000096:biosynthesis; D008969:Molecular Sequence Data; D009036:Moths; D011994:Recombinant Proteins / Q000633:toxicity; D011995:Recombination, Genetic; D014862:Wasp Venoms / Q000235:genetics* / Q000276:immunology / Q000633:toxicity; D014863:Wasps / Q000235:genetics* / Q000276:immunology
D000485:Allergens; D008543:Melanins; D011994:Recombinant Proteins; D014862:Wasp Venoms; D004247:DNA