大片形吸虫CatL1蛋白的原核表达及其特异性多克隆抗体的制备  

Prokaryotic expression of Fasciola gigantica Cathepsin L1 protein and preparation of its specific polyclonal antibody

作　　者：王乐铭 侯林静 饶国顺 梅雪芳[1] 施维[1] 王渝瑞 吴正姣 王冬英[1] 张为宇[1] WANG Leming;HOU Linjing;RAO Guoshun;MEI Xuefang;SHI Wei;WANG Yurui;WU Zhengjiao;WANG Dongying;ZHANG Weiyu(College of Animal Science and Technology,Guangxi University,Nanning 530004,China)

机构地区：[1]广西大学动物科学技术学院,南宁530004

出　　处：《黑龙江畜牧兽医》2020年第18期62-65,70,共5页Heilongjiang Animal Science And veterinary Medicine

基　　金：广西创新驱动发展专项(桂科AA17204057);国家重点研发计划项目“反刍动物肝片吸虫病早期诊断与控制技术研究”(217YFD0501202)。

摘　　要：为了制备大片形吸虫诊断抗体,试验采用原核表达制备大片形吸虫组织蛋白酶L1重组蛋白(rFgCatL1),用该蛋白免疫Balb/c小鼠制备多克隆抗体,通过间接酶联免疫吸附法(ELISA)检测抗体效价,Western-blot法检测多克隆抗体的特异性。结果表明:CatL1基因大小为981 bp,蛋白质分子质量为39.0 ku左右,与预期分子质量大小一致。制备的小鼠多克隆抗体血清效价达1∶128 000,能特异性识别大片形吸虫排泄-分泌物(ESP)抗原。说明制备的rFgCatL1多克隆抗体可作为良好的诊断抗体用于研制快速诊断试剂盒。In order to prepare a suitable antibody for the diagnosis of Fasciola gigantica(F. gigantica),F. gigantica Cathepsin L1(rFgCatL1) recombinant protein was prepared by prokaryotic expression. Balb/c mice were immunized with the prepared rFgCatL1 to produce polyclonal antibody. The antibody titer was detected by indirect ELISA. The specificity of polyclonal antibody was determined by Western-blot assay. The results showed that the size of CatL1 gene was 981 bp, and the molecular weight of the protein was about 39.0 ku, which was consistent with the theoretical molecular mass. The prepared mouse polyclonal serum titer was mensurated 1∶128 000, which could specifically recognize the F.gigantica ESP antigen. It indicated that rFgCatL1 polyclonal antibody could be used as a good diagnostic antibody to develop a rapid diagnostic kit.

关 键 词：大片形吸虫 组织蛋白酶L1 原核表达 多克隆抗体 特异性 

分 类 号：S852.735[农业科学—基础兽医学] Q78[农业科学—兽医学]