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Expression of hornet genes encoding venom allergen antigen 5 in insects
Author(s) -
Tomalski Michael D.,
King Te Piao,
Miller Lois K.
Publication year - 1993
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.940220302
Subject(s) - biology , antiserum , polyclonal antibodies , venom , gene , antigen , microbiology and biotechnology , gene expression , recombinant dna , gene product , biochemistry , immunology
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. © 1993 Wiley‐Liss, Inc.