利用核糖体展示技术筛选口蹄疫病毒单链抗体基因  

Selection of scFv Genes of Foot-and-Mouth Disease Virus Using Ribosome Display

作　　者：代鹏[1] 周广青[1] 林密[1] 冯霞[1] 马军武[1] 张晓霞[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室国家口蹄疫参考实验室甘肃省生物检测工程技术研究中心,兰州730046

出　　处：《中国生物工程杂志》2012年第7期95-101,共7页China Biotechnology

基　　金：中央级公益性科研院所基本科研业务费专项资助项目(B2F100306)

摘　　要：目的:利用核糖体展示技术筛选口蹄疫病毒特异性单链抗体基因。方法:在已构建好的核糖体展示文库的基础上,利用核糖体展示技术,经过5轮的体外转录、体外转译、亲和筛选和RT-PCR,将得到的序列进行测序分析。结果:筛选到FMDV scFv基因,且基因得到富集。结论:实验运用核糖体展示技术,以FMDV抗原和纯化的146S病毒粒子为靶标筛选到了FMDV scFv基因,将为scFv用于FMD的基础研究、免疫学研究以及为预防、治疗和诊断提供帮助,也为研制FMD的快速诊断技术奠定先前基础。Objective： To screen the specific single-chain variable Fragment （scFv） gene of Foot-and-Mouth Disease by Ribosome display. Methods ： Based on the construction of ribosome display library, five rounds of in vitro transcription, in vitro translation, affinity selection and RT-PCR constructed by ribosome display technology, then sequencing and analysis. Results： ScFv genes of FMDV were screened, and were enriched. Conclusion： Targeted by antigen of FMDV and purified 146S virus particle, FMDV scFv genes were obtained by using ribosome display technology. Not only previous basis for basic research, immunology research, prevention, treatment and diagnostic of FMD, but also for the development of rapid diagnostic techniques of FMD by using single-chain variable fragment were provided.

关 键 词：口蹄疫病毒 单链抗体 核糖体展示 

分 类 号：Q78[生物学—分子生物学]