首页> 外文期刊>World Journal of Microbiology & Biotechnology >Cry2A toxins from Bacillus thuringiensis expressed in insect cells are toxic to two lepidopteran insects
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Cry2A toxins from Bacillus thuringiensis expressed in insect cells are toxic to two lepidopteran insects

机译:苏云金芽孢杆菌在昆虫细胞中表达的Cry2A毒素对两种鳞翅目昆虫有毒

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摘要

The cry2Aa and cry2Ab genes from a Brazilian Bacillus thuringiensis strain were introduced into the genome of the baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) in order to evaluate the heterologous proteins expression in insect cells and their toxicity to different insects. The recombinant viruses (vAcCry2Aa and vSynCry2Ab) were amplified in Trichoplusia ni (BTI-Tn5B1-4) cells and used to infect Spodoptera frugiperda larvae. Total extracts from S. frugiperda infected with the recombinant viruses were analysed by SDS-PAGE, which detected the presence of polypeptides around 65 kDa. Cuboid-shaped protein crystals were observed in insect extracts by light and scanning electron microscopy. Bioassays, using the heterologous proteins showed toxicity against second instar A. gemmatalis larvae (Cry2Aa) with a LC50 of 1.03 mu g/ml and second instar S. frugiperda larvae (Cry2Ab) with a LC50 of 3.45 mu g/ml. No toxic activity was detected for Aedes aegypti and Culex quinquenfaciatus.
机译:为了评估异源蛋白在昆虫细胞中的表达及其对不同昆虫的毒性,将巴西苏云金芽胞杆菌菌株的cry2Aa和cry2Ab基因引入杆状病毒加州产白粉虫多核多角体病毒(AcMNPV)的基因组中。重组病毒(vAcCry2Aa和vSynCry2Ab)在Trichoplusia ni(BTI-Tn5B1-4)细胞中扩增,并用于感染节食夜蛾幼虫。通过SDS-PAGE分析感染重组病毒的S. frugiperda的总提取物,检测到65 kDa左右的多肽的存在。通过光镜和扫描电子显微镜在昆虫提取物中观察到长方体状的蛋白质晶体。使用异源蛋白质的生物测定法显示出对第二龄A. gemmatalis幼虫(Cry2Aa)的LC50为1.03μg / ml和第二龄S. frugiperda幼虫(Cry2Ab)的LC50为3.45μg / ml的毒性。没有检测到埃及伊蚊和库克斯库克斯昆卡西亚图斯的毒性活性。

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