口蹄疫病毒单链抗体基因的构建及其推导蛋白三维结构的分子模拟  

Construction of single chain antibody gene of FMDV and homology modeling of 3D structure of its deduced protein

作　　者：刘涛[1] 周广青[1] 马军武[1] 林密[1] 祁淑芸[1] 冯霞[1] 马明仁[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农业部畜禽病毒学重点开放实验室,甘肃兰州730046

出　　处：《中国兽医科学》2009年第12期1035-1040,共6页Chinese Veterinary Science

基　　金：国家"十一五"科技支撑计划项目(2006BAD06A10);中央级公益性科研院所基本科研业务费专项(BRF090305)

摘　　要：用口蹄疫病毒(FMDV)Asia1/JS/China/2005株灭活抗原免疫家兔,从兔脾细胞中提取RNA作为模板,通过PCR及重叠PCR方法构建出FMDV的单链抗体基因。测序结果显示,序列中有重链可变区(VH)及轻链可变区(VL)基因,经Blast比较,发现VH、VL的相似性最高分别可达81.0%和93.0%,且在序列中有连接VH及VL的柔性接头(Gly4Ser)3。用EXPASY软件包预测了推导蛋白的特性。运用Swiss-pdb viewer软件的SWISS-Model处理器对构建的单链抗体基因推导的蛋白序列进行了三维结构的分子模拟。拉马钱德兰图证明,所模拟的单链抗体的三维结构较为合理。The RNA was extracted from the splenocyte of rabbits immunized with the inactivated antigen from foot-and-mouth disease virus（FMDV） Asial/JS/China/2005 strain. The heavy variable chain VH gene and the light variable chain VL gene were amplified. The single chain antibody（ScFv） gene was constructed by over-lapping PCR. The sequence was analyzed by Blast from the GenBank database. The amplified VH and VL genes shared 81.0G and 93.0G identities with those in GenBank,respectively. And the linker（Gly4Ser）a was in the sequence of ScFv gene. The properties of protein from SeFv gene were analyzed. The homology modeling of 3D structure of the protein was done by SWISS-Model of Swiss-pdb viewer. The Ramachandran Plot showed that the structure of modeled ScFv was reasonable.

关 键 词：口蹄疫病毒 单链抗体 重叠聚合酶链反应 序列分析 三维结构 分子模拟 

分 类 号：S852.659.6[农业科学—基础兽医学]