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A recombinant fusion protein containing a spider toxin specific for the insect voltage-gated sodium ion channel shows oral toxicity towards insects of different orders

Yang, S.; Pyati, P.; Fitches, E.; Gatehouse, J.A.

A recombinant fusion protein containing a spider toxin specific for the insect voltage-gated sodium ion channel shows oral toxicity towards insects of different orders Thumbnail


Authors

S. Yang

P. Pyati

J.A. Gatehouse



Abstract

Recombinant fusion protein technology allows specific insecticidal protein and peptide toxins to display activity in orally-delivered biopesticides. The spider venom peptide δ-amaurobitoxin-PI1a, which targets insect voltage-gated sodium channels, was fused to the “carrier” snowdrop lectin (GNA) to confer oral toxicity. The toxin itself (PI1a) and an amaurobitoxin/GNA fusion protein (PI1a/GNA) were produced using the yeast Pichia pastoris as expression host. Although both proteins caused mortality when injected into cabbage moth (Mamestra brassicae) larvae, the PI1a/GNA fusion was approximately 6 times as effective as recombinant PI1a on a molar basis. PI1a alone was not orally active against cabbage moth larvae, but a single 30 μg dose of the PI1a/GNA fusion protein caused 100% larval mortality within 6 days when fed to 3rd instar larvae, and caused significant reductions in survival, growth and feeding in 4th – 6th instar larvae. Transport of fusion protein from gut contents to the haemolymph of cabbage moth larvae, and binding to the nerve chord, was shown by Western blotting. The PI1a/GNA fusion protein also caused mortality when delivered orally to dipteran (Musca domestica; housefly) and hemipteran (Acyrthosiphon pisum; pea aphid) insects, making it a promising candidate for development as a biopesticide.

Citation

Yang, S., Pyati, P., Fitches, E., & Gatehouse, J. (2014). A recombinant fusion protein containing a spider toxin specific for the insect voltage-gated sodium ion channel shows oral toxicity towards insects of different orders. Insect Biochemistry and Molecular Biology, 47, 1-11. https://doi.org/10.1016/j.ibmb.2014.01.007

Journal Article Type Article
Acceptance Date Jan 21, 2014
Publication Date Apr 1, 2014
Deposit Date Mar 2, 2015
Publicly Available Date Mar 29, 2024
Journal Insect Biochemistry and Molecular Biology
Print ISSN 0965-1748
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 47
Pages 1-11
DOI https://doi.org/10.1016/j.ibmb.2014.01.007
Keywords Neurotoxin, Insecticide, Recombinant protein expression system, Protein transport, Crop protection, Biotechnology.

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