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Insect Immune Activation by Recombinant Apolipophorin‐III and Its Application to Pest Control in Hyphantria cunea
Author(s) -
KANG YoungJin,
KIM HongJa,
SEO SookJae
Publication year - 2003
Publication title -
entomological research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.421
H-Index - 20
eISSN - 1748-5967
pISSN - 1738-2297
DOI - 10.1111/j.1748-5967.2003.tb00071.x
Subject(s) - hemolymph , hyphantria , biology , galleria mellonella , western blot , microbiology and biotechnology , biochemistry , botany , gene , larva , virulence
ABSTRACT Apolipophorin‐III is a hemolymph protein whose function is to facilitate lipid transport in an aqueous medium. Recently, apolipophorin‐III in Galleria mellonella larvae showed to play an unexpected role in insect immune activation. We identified the cDNA sequence of Hyphantria cunea apolipophorin‐III by oligonucleotide‐primed amplification, and 5′‐ and 3′‐RACE PCR. Since H. cunea has an unusually low level of apolipophorin‐III in the hemolymph, a recombinant apolipophorin‐III was overexpressed using a baculovirus expression system to investigate its biological activity. Recombinant apolipophorin‐III and E. coli were injected into the hemocoel of last instar larvae, and the changes in apolipophorin‐III in hemolymph was determined by Western blot. Injection of apolipophorin‐III induced a slight increase of apolipophorin‐III in the recipients'hemolymph. While E. coli injection led to remarkably increased concentration of apolipophorin‐III in hemolymph. To investigate the induction of antimicrobial peptide by the injection of recombinant apoLp‐III and E. coli , Northern blot was performed. Apolipophorin‐III injection as well as bacterial injection into the larvae showed the induction on the expression of lysozyme gene. Apolipophorin‐III is apparently related to the immune response through an unknown mechanism. The role of apolipophorin‐III in insect immunity should be related to the activation of transcription factor of antimicrobial peptide.