【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。更多还原

【Abstract】 The RNA was extracted from the splenocyte of rabbits immunized with the inactivated antigen from foot-and-mouth disease virus(FMDV) Asia1/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 81.0% and 93.0% identities with those in GenBank,respectively.And the linker(Gly4Ser)3 was in the sequence of ScFv gene.The properties of protein from ScFv 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.更多还原