| 抗牛结核分枝杆菌VHH抗体T7噬菌体库的构建与筛选 |
| |
| 引用本文： | 杨艳丽,张齐,张星星,许龙,黄新,吴桐忠,韩猛立,钟发刚,张小莺.抗牛结核分枝杆菌VHH抗体T7噬菌体库的构建与筛选[J].中国兽医学报,2019(10):1987-1993. |
| |
| 作者姓名： | 杨艳丽 张齐 张星星 许龙 黄新 吴桐忠 韩猛立 钟发刚 张小莺 |
| |
| 作者单位： | 1.新疆农业大学动物医学院;2.新疆农垦科学院省部共建绵羊遗传改良与健康养殖国家重点实验室;3.Centre of Molecular and Environmental Biology;4.University of Minho |
| |
| 基金项目： | 乌鲁木齐市科学技术计划资助项目(P161310001) |
| |
| 摘 要： | 为获得抗牛结核分枝杆菌的VHH纳米抗体,本研究利用牛结核分枝杆菌Ag85B蛋白免疫双峰驼,免疫效价达到1∶8 000后采外周血分离淋巴细胞,提取RNA反转录成cDNA,利用巢式PCR方法扩增出编码单域抗体VHH的400 bp基因片段。将目的片段用限制性内切酶EcoRⅠ和HindⅢ双酶切,与T7噬菌体载体臂T7Select 10-3RI Arms连接,再通过体外重新包装含目的片段的噬菌体。对建立的VHH噬菌体抗体库进行鉴定,原始文库库容为5.7×10~8 PFU,库阳性率为85.7%。通过4轮生物亲和淘选,筛选并表达出2株高特异性的VHH抗体,均对牛结核分枝杆菌Ag85B蛋白具有特异性。本研究为进一步探讨VHH抗体在牛结核病的诊断和治疗奠定了基础。
|
| 关 键 词： | 牛结核分枝杆菌 Ag85B蛋白 VHH抗体 T7噬菌体展示 |
Construction and selection of VHH antibody against bovine M. tuberculosis using T7 phage display |
| |
| YANG Yan-li,ZHANG Qi,ZHANG Xing-xing,XU Long,HUANG Xin,WU Tong-zhong,HAN Meng-li,ZHONG Fa-gang,ZHANG Xiao-ying.Construction and selection of VHH antibody against bovine M. tuberculosis using T7 phage display[J].Chinese Journal of Veterinary Science,2019(10):1987-1993. |
| |
| Authors: | YANG Yan-li ZHANG Qi ZHANG Xing-xing XU Long HUANG Xin WU Tong-zhong HAN Meng-li ZHONG Fa-gang ZHANG Xiao-ying |
| |
| Institution: | (College of Veterinary Medicine ,Xin-jiang Agricultural University ,Urumqi 830000, China;State Key Laboratory for Sheep Genetic Improvement and Healthy Production ,Xinjiang Academy of Agricultural and Reclamation Science , Shihezi, Xinjiang 832000 ,China;College of Veterinary Medicine,Northwest A & F University,Yangling,Shaanxi 712100 ,China) |
| |
| Abstract: | YANG Yan-li;ZHANG Qi;ZHANG Xing-xing;XU Long;HUANG Xin;WU Tong-zhong;HAN Meng-li;ZHONG Fa-gang;ZHANG Xiao-ying(College of Veterinary Medicine ,Xin-jiang Agricultural University ,Urumqi 830000, China;State Key Laboratory for Sheep Genetic Improvement and Healthy Production ,Xinjiang Academy of Agricultural and Reclamation Science , Shihezi, Xinjiang 832000 ,China;College of Veterinary Medicine,Northwest A & F University,Yangling,Shaanxi 712100 ,China) |
| |
| Keywords: | bovine Mycobacterium tuberculosis protein Ag85B variable domain of heavy-chain antibodies ( VHH ) T7 phage display |
| 本文献已被 CNKI 维普 等数据库收录！ |
|
