猪蛔虫卵可溶性抗原单链抗体库的构建及初步筛选.pdfVIP

贵州农业科学 2011 , 39(12):178--181 Guizhou Agricultural Sciences [文章编号]1001-3601(2011)12-0769-0178-04 猪虫回虫卵可溶性抗原单链抗体库的构建及初步筛选 何光志1 ，周颖2 ，安传伟3 ，张太华2 ，熊文康2 ，崔小东2 ，罗成松2 ，谢波2 (1.贵阳中医学院基础部，贵州贵阳 550002; 2. 思南县动物疾病预防控制中心，贵州思南 565100; 3. 贵阳 中医学院时珍学院 2010 级中西结合临床，贵州贵阳 550002) [摘 要]为进一步研制抗猪拥虫的高特异性、高亲和力的基因工程抗体奠定基础，以猪拥虫未成熟虫 卵可溶性抗原免疫 BALB/C 小鼠，提取小鼠脾脏总 RNA，以 RT-PCR 技术分别扩增出 H 链和 L 链可变区 基因，以 SOE-PCR 法将 VH 和 VL 片段随机拼接成 ScFv 片段，然后将 ScFv 片段克隆至 pCANTAB-5E 载 体，并转化至感受态 TGl 菌，经辅助噬菌体 M13K07 拯救，构建猪捆虫卵可溶性抗原噬菌体展示羊链抗体 库，结果从 20 个噬菌体克隆中筛选到 15 个阳性克隆。 [关键词]拥虫;可溶性抗原;羊链抗体库;构建与筛选 [中图分类号] S858.28 [文献标识码]A Construction and Screening of Single-Chain Antibody Library of Eggs of Soluble Antigen from Ascαris suum 1 2 3 2 HE Guang-zhi , ZHOU Ying , AN Chuan-wei , ZHANG Tai-hua , 2 1 2 XIONG Wen-kang , CUI Xiao-dong , LUO Cheng-song , XIE B02 (1. Guiyang College 01 Traditional Chinese Medicine , Guiyang , Guizhou 550002; 2. Sinan Prevention and Regulation Center 01 Animal Deasease , Sinan , Guizhou 565100; 3.2010 Graduates Chinese and Western Clinical Medicine , Shizhen College , Guiyang College 01 Traditional Chinese Medicine , Guiyang , Guizhou 550002 , China) Abstract: To lay a solid foundation for preparation of mice engineering antibody to A. suum with higher affinity , mice were injected with the recombinant protein eggs of soluble antigen from A. suum mixed with FCA. Total RNA was extracted from the spleen cell of the immune mice followed RT-PC R. The heavy-chain and light-chain variable region genes (VH and VL) repertoire of immunoglobulin were amplified respectively by PCR. Then they we