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首页> 外文期刊>Pesticide Biochemistry and Physiology >Fluorescent analyses of Bacillus thuringiensis Cry1Fa and Cry1Ab toxin binding sites on brush border membrane vesicles of Ostrinia nubilalis (Hubner), Diatraea grandiosella (Dyar), and Helicoverpa zea (Boddie) larvae
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Fluorescent analyses of Bacillus thuringiensis Cry1Fa and Cry1Ab toxin binding sites on brush border membrane vesicles of Ostrinia nubilalis (Hubner), Diatraea grandiosella (Dyar), and Helicoverpa zea (Boddie) larvae

机译：芽孢杆菌的荧光分析芽孢杆菌Cry1Fa和Cry1ab毒素瘤瘤卷眼膜囊泡囊泡囊泡（Hubner），DiaTraea Grandioslella（Dyat）和Helicoverpa Zea（Boddie）幼虫

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摘要

Bacillus thuringiensis (Bt) Cry1Fa and Cry1Ab proteins are important Cry toxins due to their high, selective toxicity against a number of lepidopteran species, including important pests of corn and cotton. Competition binding assays are a classical tool for investigating Cry toxin interactions with target pest insects. We developed a fluorescence-based binding assay and assessed Cry1Fa and Cry1Ab toxin binding to brush border membrane preparations from lepidopteran corn pests including Ostrinia nubilalis (European corn borer, ECB), Diatraea grandiosella (south western corn borer, SWCB), and Helicoverpa zea (corn earworm, CEW). Homologous and heterologous competition binding assays with fluorophore-(Alexa488)-labeled Cry1Fa toxin showed that Cry1Fa shares binding site(s) with Cry1Ab toxin in ECB, and SWCB for which Cry1Ab has higher affinity than Cry1Fa. Apart from the shared binding sites, Cry1Ab and Cry1Fa bind an additional site(s) in ECB and SWCB. In CEW, Cry1Fa and Cry1Ab each, has a high affinity binding site(s), which binds the heterologous toxin with low affinity. The Cry1Ab-Cry1Fa toxin binding models for ECB, SWCB and CEW based on our results are considered in the context of what is known about acquired cross-resistance against Cry1Ab and Cry1Fa toxins.
机译：Bacillus thuringiensis（BT）Cry1Fa和Cry1ab蛋白是重要的毒毒素，因为它们的高度选择性毒性对许多鳞片类物种，包括玉米和棉花的重要害虫。竞争结合测定是一种古典工具，用于研究与靶害虫昆虫的哭泣毒素相互作用。我们开发了一种基于荧光的结合测定和评估的Cry1Fa和Cry1ab毒素结合，从鳞翅目玉米害虫（欧洲玉米螟，欧洲央行），DiaTraea GrandioSella（南食玉米螟，SWCB）和Helicoverpa Zea（玉米耳虫，克服）。用荧光团（Alexa488） - 标记的Cry1FA毒素的同源和异源竞争结合测定结果表明，Cry1Fa在欧洲央行中用Cry1ab毒素共享结合位点，并且Cry1ab的SWCB比Cry1Fa具有更高的亲和力。除了共享绑定站点，cry1ab和cry1fa在ecb和swcb中绑定附加站点。在CEW中，Cry1Fa和Cry1ab各自具有高亲和力结合位点，其结合具有低亲和力的异源毒素。基于我们的结果的Cry1ab-Cry1FA毒素绑定模型用于欧洲央行，SWCB和CEW的基础是在所知的关于对Cry1ab和Cry1FA毒素的跨性抗性的知名度的上下文中。

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