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Computational and biological characterization of fusion proteins of two insecticidal proteins for control of insect pests

机译:用于控制害虫的两种杀虫蛋白融合蛋白的计算和生物学表征

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摘要

Sucking pests pose a serious agricultural challenge, as available transgenic technologies such as Bacillus thuringiensis crystal toxins (Bt) are not effective against them. One approach is to produce fusion protein toxins for the control of these pests. Two protein toxins, Hvt (ω-atracotoxin from Hadronyche versuta) and onion leaf lectin, were translationally fused to evaluate the negative effects of fusion proteins on Phenacoccus solenopsis (mealybug), a phloem-feeding insect pest. Hvt was cloned both N-terminally (HL) and then C-terminally (LH) in the fusion protein constructs, which were expressed transiently in Nicotiana tabacum using a Potato Virus X (PVX) vector. The HL fusion protein was found to be more effective against P. solenopsis, with an 83% mortality rate, as compared to the LH protein, which caused 65% mortality. Hvt and lectin alone caused 42% and 45%, respectively, under the same conditions. Computational studies of both fusion proteins showed that the HL protein is more stable than the LH protein. Together, these results demonstrate that translational fusion of two insecticidal proteins improved the insecticidal activity relative to each protein individually and could be expressed in transgenic plants for effective control of sucking pests.
机译:由于诸如苏云金芽孢杆菌晶体毒素(Bt)等可用的转基因技术对它们无效,因此,吸吮害虫构成了严重的农业挑战。一种方法是产生融合蛋白毒素以控制这些害虫。将两种蛋白毒素Hvt(来自Hadronyche versuta的ω-atracotoxin)和洋葱叶凝集素进行翻译融合,以评估融合蛋白对食韧皮部飞虫(Phenacoccus solenopsis)(mealybug)的负面影响。将Hvt克隆到融合蛋白构建体的N端(HL)和C端(LH),然后使用马铃薯病毒X(PVX)载体在烟草中瞬时表达。发现HL融合蛋白更有效地抵抗了肺炎疫霉,与导致65%死亡率的LH蛋白相比,其死亡率为83%。在相同条件下,单独的Hvt和凝集素分别导致42%和45%。两种融合蛋白的计算研究表明,HL蛋白比LH蛋白更稳定。总之,这些结果表明,两种杀虫蛋白的翻译融合相对于每种蛋白分​​别提高了杀虫活性,并且可以在转基因植物中表达以有效控制吸吮性害虫。

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