昆虫病原细菌Cip蛋白多克隆抗体的制备  

Preparation of Polyclonal Antibody against Crystalline Inclusion Protein of Entomopathogenic Bacterium

作　　者：游娟[1] 黄建林 曹莉[3] 韩日畴[3] 

机构地区：[1]广东药学院生物化学与分子生物学系,广东广州510006 [2]广州计量检测技术研究院,广东广州510030 [3]广东省昆虫研究所,广东广州510260

出　　处：《河南农业科学》2013年第8期62-66,共5页Journal of Henan Agricultural Sciences

基　　金：广东省自然科学基金项目(036692)

摘　　要：昆虫病原发光杆菌属(Photorhabdus)细菌产生2种胞内晶体蛋白CipA和CipB,为其共生线虫的生长发育提供营养。基于Cip蛋白已在大肠杆菌原核表达系统中得到稳定且高水平表达,以纯化的重组Cip蛋白为抗原,免疫新西兰大耳兔制备多克隆抗体,ELISA测定2种蛋白抗血清效价,均可达到1∶6 400。Western blot分析显示,制备的抗体均可特异识别发光杆菌的胞内晶体蛋白。Cip蛋白多克隆抗体的制备为建立该类蛋白的免疫学检测方法奠定了基础。Entomopathogenic Photorhabdus bacteria produce two types of intracellular crystalline inclusion proteins,CipA and CipB,as the nutrient resource for the symbiotic nematode.Based on constructed high-level prokaryotic expression system,recombinant Cip proteins were purified with a warm extract method and used as the antigens to immune New Zealand rabbits,respectively.The polyclonal antibodies with the titers of 1：6400 were raised and monitored by indirect enzyme-linked immunosorbent assay （ELISA）.Western blot assay confirmed that the antibodies could specifically recognize the inclusion proteins in Photorhabdus bacteria.Preparation of Cip polyclonal antibodies will be helpful to develop the immunological assay method of these proteins.

关 键 词：昆虫病原细菌 发光杆菌属 胞内晶体蛋白 多克隆抗体 制备 

分 类 号：S476.11[农业科学—农业昆虫与害虫防治]