小菜蛾杀虫抗体的对靶设计及验证  

Target design and verification of insecticidal antibody against Plutella xylostella

作　　者：谢雅晶 杨立应[1] 胡晓丹 徐重新 张霄 高美静[2] 卢莉娜[2] 仲建锋 朱庆 刘媛[1,2] 刘贤金 XIE Yajing;YANG Liying;HU Xiaodan;XU Chongxin;ZHANG Xiao;GAO Meijing;LU Lina;ZHONG Jianfeng;ZHU Qing;LIU Yuan;LIU Xianjin(School of Food and Biological Engineering,Jiangsu University,Zhenjiang 212013,China;Key Laboratory of Food Quality and Safety of Jiangsu Province-State Key Laboratory Breeding Base of Ministry of Science and Technology/Key Laboratory of Control Technology and Standard for Agro-product Safety and Quality,Ministry of Agriculture and Rural Affairs/Key Laboratory for Agro-product Safety Risk Evaluation(Nanjing),Ministry of Agriculture and Rural Affairs/Institute of Food Safety and Nutrition,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;Jiangsu Vocational College of Agriculture and Forestry,Zhenjiang 212400,China)

机构地区：[1]江苏大学食品与生物工程学院,江苏镇江212013 [2]江苏省食品质量安全重点实验室-省部共建国家重点实验室培育基地/农业农村部农产品质量安全控制技术与标准重点实验室/农业农村部农产品质量安全风险评估实验室<南京>/江苏省农业科学院农产品质量安全与营养研究所,江苏南京210014 [3]江苏农林职业技术学院,江苏镇江212400

出　　处：《江苏农业学报》2024年第7期1212-1219,共8页Jiangsu Journal of Agricultural Sciences

基　　金：国家自然科学基金青年基金项目(31301703);江苏省农业科技自主创新基金项目[CX(22)1009];江苏现代农业产业技术体系建设专项[JATS(2023)403];江苏省自然科学基金面上项目(BK20231384)

摘　　要：本研究拟通过模拟Cry毒素创制新型杀虫蛋白质用于小菜蛾防治,主要利用三维结构模拟及分子对接技术,以前期获得的抗Cry1Ab抗体为模板设计2个基因工程抗体(GEAb)。其中,具有杀虫活性的GEAb-GGCC与小菜蛾中肠刷状边缘膜囊泡(BBMV)具有较高结合活性,并与Cry1A和Cry1B具有重叠的BBMV结合位点。BBMV免疫沉淀分析鉴定结果显示,与GEAb-GGCC结合的中肠蛋白质包括氨肽酶N(APN)、V-ATP酶B亚基和polycalin。由于GEAb-GGCC缺乏Cry1A类蛋白质中负责成孔的α-螺旋结构,推测GEAb-GGCC或通过结合小菜蛾中肠受体激活下游信号通路,引起中肠损伤,导致虫体死亡。To generate new insecticidal proteins against Plutella xylostella by mimicking Cry toxins,two genetically engineered antibodies(GEAb)were designed by using previously obtained anti-Cry1Ab idiotypic-antibody as template,mainly by three-dimensional structure modeling and molecular docking technology.The insecticidal GEAb-GGCC of GEAb was found to show high affinity to P.xylostella brush border membrane vesicles(BBMV),and had superposed BBMV binding sites with Cry1A and Cry1B.Identification results of BBMV immunoprecipitation analysis showed that the midgut proteins bonded with GEAb-GGCC included aminopeptidases N(APN),B subunit of V-ATPase and polycalin.Because the GEAb-GGCC is lack ofα-helix structure which is responsible for pore forming in Cry1A proteins,it is speculated that GEAb-GGCC may activate the downstream signal pathway by binding with the midgut receptors to cause midgut injury,which leads to the death of P.xylostella larva.

关 键 词：对靶设计 杀虫基因工程抗体 分子对接 小菜蛾 Cry毒素 

分 类 号：Q816[生物学—生物工程]