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.
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.
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DOI：
10.1016/j.ibmb.2014.01.007
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发表时间：
2014-04
影响因子：
3.8
通讯作者：
Gatehouse, John A.
中科院分区：
文献类型：
--
作者：
Yang, Sheng;Pyati, Prashant;Fitches, Elaine;Gatehouse, John A.
关键词：
重组融合蛋白技术允许特定的杀虫蛋白和肽毒素在口服递送的生物农药中显示活性。蜘蛛毒肽δ-amaurobitoxin-PI1a靶向昆虫电压门控钠通道，与"载体"雪花莲凝集素（GNA）融合以赋予口服毒性。利用毕赤酵母作为表达宿主，生产毒素本身（PI1a）和amaurobitoxin/GNA融合蛋白（PI1a/GNA）。虽然这两种蛋白质注射到甘蓝蛾（Mamestra brassicae）幼虫时引起死亡率，PI1a/GNA融合是约6倍的有效重组PI1a的摩尔基础上。单独的PI1a对甘蓝蛾幼虫没有口服活性，但是当饲喂3龄幼虫时，单次30 μ g剂量的PI1a/GNA融合蛋白在6天内引起100%的幼虫死亡率，并且引起4 - 6龄幼虫的存活、生长和摄食的显著降低。蛋白质印迹法显示融合蛋白从消化道内容物运输到小菜蛾幼虫血淋巴，并结合到神经索。PI1a/GNA融合蛋白在口服递送给双翅目（家蝇;家蝇）和半翅目（豌豆蚜;豌豆蚜）昆虫时也引起死亡，使其成为开发为生物农药的有希望的候选者。蜘蛛毒素Pl1a，对昆虫钠离子通道特异性，单独表达，并与雪花莲凝集素（GNA）融合。重组毒素和融合蛋白对家蝇成虫和菜蛾幼虫均有杀虫活性。只有融合蛋白对家蝇成虫和菜蛾幼虫具有口服毒性。与其组分相比，融合蛋白显示出增强的对蚜虫的口服毒性。Pl1a/GNA融合蛋白具有作为生物农药开发的前景。
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. The spider toxin Pl1a, specific for the insect sodium channel, was expressed alone and as a fusion with snowdrop lectin (GNA). Both recombinant toxin and fusion protein were insecticidal to housefly adults and cabbage moth larvae when injected. Only the fusion protein was orally toxic to housefly adults and cabbage moth larvae. The fusion protein showed enhanced oral toxicity towards aphids compared to its components. The Pl1a/GNA fusion protein has promise for development as a biopesticide.
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影响因子：
3.8
作者：
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通讯作者：
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3.5
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