摘要
Vip3A类杀虫蛋白是一类新型杀虫蛋白,在氨基酸序列上与研究较为深入的Cry蛋白没有同源性,且昆虫受体结合位点与Cry蛋白也没有竞争关系。利用同源重组技术针对对甜菜夜蛾具有高活性的Vip3Aa39与无活性的Vip3Ad开展研究,成功构建了三个vip3AaAdAa型嵌合基因,将其转入大肠杆菌中表达嵌合蛋白Vip3AaAdAa1、Vip3AaAdAa2、Vip3AaAdAa3,并将这三种嵌合蛋白对甜菜夜蛾进行了杀虫活性测定。研究表明,三种嵌合蛋白对甜菜夜蛾具有杀虫活性,Vip3AaAdAa3蛋白的LC_(50)值比Vip3Aa39增加了1.4倍,Vip3AaAdAa1蛋白的LC_(50)值比Vip3Aa39增加了2.7倍,嵌合蛋白Vip3AaAdAa1和Vip3AaAdAa3的杀虫活性均低于Vip3Aa39;Vip3AaAdAa2的杀虫活性与Vip3Aa39差异不显著。
Vip3A-type insecticidal proteins are a new class of insecticidal proteins. There is no homology in amino acid sequences with well-studied protein Cry,and the insecticidal sites are not competitive with insect receptor binding sites of Cry proteins. Homologous recombinant technique was used to study the insecticidal activity of Vip3Aa39 and inactive Vip3Ad against Spodoptera exigua. Three vip3AaAdAa chimeric genes were successfully created and were transformed into Escherichia coli to express chimeric protein Vip3AaAda1,Vip3AaAda2,and Vip3AaAda3. The insecticidal activities of these 3 chimeric proteins against S. exigua were determined. The results showed that the 3 chimeric proteins presented high insecticidal activities against S. exigua,the LC_(50) of Vip3AaAda3 protein was 1.4 times higher than that of Vip3Aa39,and the LC_(50) of Vip3AaAda1 protein was 2.7 times higher than that of Vip3Aa39. The insecticidal activities of chimeric proteins Vip3AaAda1 and Vip3AaAda3 were lower than that of Vip3Aa39,and the insecticidal activity of Vip3AaAdAa2 was not significantly different from that of Vip3Aa39.
引文
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