【摘要】 用ＰＣＲ方法以分泌抗甜菜坏死黄脉病毒（ＢＮＹＶＶ）单克隆抗体的杂交瘤细胞的基因组为模板，扩增了编码ＢＮＹＶＶ单抗的重链可变区（ＶＨ）基因。测序表明，该ＶＨ序列属于小鼠Ⅱ（Ａ）亚类，全长为３６０ｂｐ，编码１２０个氨基酸。将其和先前克隆的轻链基因分别插入到一个含有连接ＶＨ和ＶＬ基因的连接序列的质粒之中，构建成单链抗体（ｓｃＦｖ）基因的表达载体ｐＴＣｓｃＦｖ。将质粒在大肠杆菌中表达，ＥＬＩＳＡ法检测出表达产物有与ＢＮＹＶＶ抗原结合的活性。更多还原

【Abstract】 The heavy chains variable region gene (VH) of monoclonal antibody against beet necrotic yellow vein virus (BNYVV) was amplified from total DNA extracted from anti-BNYVV hybridoma cells by PCR. Sequencing showed that the VH belongs to mouse subgroup Ⅱ(A) and contains 360bp, which code one hundred and twenty amino acids. The VH and VL genes were inserted into a plasmid which contains a linker sequence for constructing scFv gene. The new vector named pTC scFV. The scFv was produced in Escherichia coli and appeared binding activity with BNYVV antigen by ELISA method.更多还原