用基因融合技术进行杀蚊幼毒蛋白基因的研究Ⅱ.蚊幼虫肠壁细胞膜抗体基因的克隆

STUDIES ON THE LARVICIDAL TOXIC PROTEIN GENE WITH THE TECHNIQUE OF GENE FUSION Ⅱ.CLONING OF ANTIBODY GENE OF MOSQUITO LARVA GUT CELL MEMBRANE

为了提高 Ｂ．ｓ．对蚊幼虫肠壁细胞膜的识别与结合力从而提高其杀蚊活性，本实验解剖了万余只中华按蚊幼虫肠道，制备了按蚊幼虫肠壁膜泡抗原及其鼠抗血清。抗原蛋白组分分析表明膜泡的纯度较高，而抗血清实验则显示出抗体组分与肠壁的结合作用。利用 Ｐ Ｃ Ｒ 技术成功扩增了小鼠抗体重链区基因，借助 Ｔ４ Ｄ Ｎ Ａ 聚合酶的修饰，使抗体基因得以成功克隆，为研制抗体基因与毒蛋白基因融合编码的“生物导弹”打下了基础。

It has been known that the larvicidal activity depends on its binding affinity with larva gut cell membrane. In this study,we dissected more than ten thousands of Anopheles sinensis larva guts and prepared their cell membrane vesicle antigen and mouse antisera. The differences of prorein fraction between membrane vesicle and homogenate indicated the purity of the membrane preparation. While, the effect of mouse antisera against larva was reduced with larva age. This suggested that the antibody components had binding activity in early larva gut cell membrane. Besides,we succeeded in amplifying the antibody gene with the help of PCR and getting its clone with the decoration of T 4DNA polymerase. This study was beneficial to the research of "biomissile" encoded with fusion genes of the antibody gene and the toxic protein gene.