安徽农业科学2011，Vol.39Issue(20)：12223-12224,2.
鸡新城疫病毒HN基因片段的原核表达及其单抗制备
Prokaryotic Expression of Chicken NDV-HN Gene Fragment and Preparation of Its Monoclonal Antibody
摘要
Abstract
[Objective] The aim was to study characteristics of the NDV-HN protein epitope in the animal immune response. [ Method] A pair of specific primers was designed according to the HN gene sequence and multiple clone sites of vector PGEX-4T-1; and then PCR amplification was conducted by using a recombinant plasmid as template. The amplified fragment was inserted into pGEX-4T-l to construct a new recombi-nant plasmid. After the recombinant plasmid was identified, it was expressed in prokaryotic cells. The monoclonal antibody of NDV-HN was also_ prepared and identified. [Result] PCR result showed that a DNA fragment of 850 bp in length was amplified as expected; identification of recombinant plasmid revealed that the target gene was successfully constructed into the plasmid pcGEX-4T-HN23, by which a fusion protein with 57kD was expressed. The spleen cells of the mouse immunized with the purified fusion protein were fused with SP2/0 cells. After ELISA screening and subcloning for three times,two strains which could generate stably more than 20 times and produce antibody to HN protein were obtained. [ Conclusion] The resulting monoclonal antibody against NDV-HN protein provided important materials for investigating the role of HN in pathopoiesis process and its specific diagnosis.关键词
新城疫病毒/HN/原核表达/单抗Key words
NDV/ HN/ Prokaryotic expression/ Monoclonal antibody分类
农业科技引用本文复制引用
王贤,陈芳芳,余为一..鸡新城疫病毒HN基因片段的原核表达及其单抗制备[J].安徽农业科学,2011,39(20):12223-12224,2.基金项目
国家自然基金项目(30671537)资助. （30671537）
