Creation of proteinaceous insecticide from Bacuillus thuringlensis δ-endotoxin and single chain variable fragment

以苏云金芽孢杆菌δ内毒素和单克隆抗体单链可变片段为原料，制备了蛋白型杀虫剂。构建了针对天牛中阵风膜蛋白的单克隆抗体。利用该抗体制备了黄檀单链可变片段。通过基因工程将单链可变片段与苏云金芽孢杆菌δ内毒素连接起来，证实了嵌合体蛋白的结合能力。今后，我们将对这些蛋白的杀虫活性进行评价。另一方面，通过对家蚕中肠Cry1Aa δ-内毒素受体的研究，了解了δ-内毒素靶分子活性的不可缺少性，因为选择合适的分子作为单链变量的靶分子对嵌合体蛋白的活性至关重要。结果表明，Cry1Aa和Cry9Da利用氨基肽酶N作为家蚕中肠细胞的受体。Cry1Aa结合到由约60个氨基酸残基组成的氨基肽酶N的N端区域。

Creation of proteinaceous insecticide was challenged using Bacillus thuringiensis δ-endotoxin and single chain variable fragment of monoclonal antibody as materials.Monoclonal antibody was constructed against membrane protein of the midgust from longicorn beetle. Acalolepta luxuriosa and single chain variable fragment was made from the antibody. single chain variable fragment and Bacillus thuringiensis δ-endotoxin was connected by geen engineering and binding capability of the chimera-proteins were confirmed. From now on we will assess the insecticidal activity of the proteins using Acalolepte luxuriosa.On the other hand, receptor of the Cry1Aa δ-endotoxin on the midgut of the silkworm, Bombyx mori was researched to know indispensable nature for the activity of the target molecule of the δ-endotoxin since it seems most important for the activity of the chimera-protein to select appropriate molecules as targets of the single chain variable. As a result, it was shown that Cry1Aa and Cry9Da utilize aminopeptidase N as a receptor on the midgut cells of the silkworm. Cry1Aa bound to N terminal region of aminopeptidase N consisting of about 60 amino acid residues.