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[摘要]
将融合基因mCry1AbVip3A构建原核表达载体pET30A-mCry1AbVip3A,转入大肠杆菌 BL21(DE3),通过优化诱导条件获得mCry1AbVip3A蛋白。室内生测mCry1AbVip3A蛋白对亚洲玉米螟和草地贪夜蛾的杀虫效果,发现饲喂终浓度为128.44 ng/ml的mCry1AbVip3A蛋白的亚洲玉米螟1龄幼虫和草地贪夜蛾1龄、2龄幼虫存活率均为0;草地贪夜蛾3龄和4龄幼虫的存活率分别为23.33%和36.67%。结果表明,mCry1AbVip3A蛋白对亚洲玉米螟1龄幼虫的杀虫效果与Cry1Ab-ma蛋白无显著差异,但显著高于Vip3A蛋白;mCry1AbVip3A蛋白对草地贪夜蛾3龄和4龄幼虫的杀虫效果与Vip3A蛋白无显著差异,但显著高于Cry1Ab-ma蛋白。构建植物过表达载体pTF101.1-mCry1AbVip3A进行玉米遗传转化并获得转基因事件,为转基因抗亚洲玉米螟和草地贪夜蛾玉米育种提供新材料。
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[Abstract]
The prokaryotic expression vector pET30A-mCry1AbVip3A was constructed and transferred into E. coli BL21 (DE3) for induction and expression, and mCry1AbVip3A protein was obtained by optimizing induction conditions. Lab bioassay of the insecticidal effect of mCry1AbVip3A protein on Ostrinia furnacalis and Spodoptera frugiperda was performed. The final concentration of 128.44 ng/ml of mCry1AbVip3A protein was fed to Ostrinia furnacalis and Spodoptera frugiperda. The result showed that the survival rates of the 1st instar larvae of Ostrinia furnacalis, and the 1st and 2nd instar larvae of Spodoptera frugiperda were 0; the survival rates of the 3rd and 4th instar larvae of Spodoptera frugiperda were 23.33% and 36.67%, respectively. Comparative analysis showed that the insecticidal effect of mCry1AbVip3A protein and Cry1Ab-ma protein on the first instar larvae of Ostrinia furnacalis was not significantly different, but significantly higher than the Vip3A protein; the insecticidal effect of mCry1AbVip3A on the 3rd and 4th instar larvae of Spodoptera frugiperda was not significantly different from that of Vip3A protein, but was significantly higher than that of Cry1Ab-ma protein. The plant overexpression vector pTF101.1-mCry1AbVip3A was constructed for maize genetic transformation and transgenic events were obtained, providing new materials for transgenic corn breeding resistant to Ostrinia furnacalis and Spodoptera frugiperda.
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