安徽农业科学2012，Vol.40Issue(26)：12915-12916,2.
鸡新城疫病毒F基因片段的原核表达及其抗F蛋白单克隆抗体的制备
Prokaryotic Expression of Fragment of Chicken NDV-F Gene and Preparation of Its Monoclonal Antibody
摘要
Abstract
[Objective]To produce monoclonal antibody to newcastle disease virus (NDV) fusion protein F. [Method]The recombined fragment F3O6 containing NDV-F gene was cloned and expressed and its monoclonal antibody was produced with hybridoma technique. [ Result ] Firstly, a pair of specific primers were designed according to the multi-clones position of the F gene and carrier PGEX-4T-1 and a DNA fragment of 306 bp in length was amplified from a recombinant plasmid kept in our laboratory. Then this fragment was inserted into pGEX-4T-1 to restructure the plasmid pGEX-4T-l -F306. Secondly it was transformed into a strain of E. coli( BL21) and expressed in prokaryotic cells. Then the purified fusion protein with 35. 8kDa as antigen immunized Balb/c mice. Finally the mouse spleen cells were fused with SP2/0 cells. With ELISA and by sub-cloning for three times, we obtained two strains cells producing antibody to F protein. After cell passage for 10 times, the antibody titers kept a-bove 1:5 124 h in cell supernatant and 1:64 000 in the ascites. [Conclusion]Two strains of hybridoma cells were obtained, which could stably produce the monoclonal antibody against NDV-F protein.关键词
新城疫病毒/F蛋白/原核表达/单克隆抗体Key words
NDV/F protein/Prokaryotic expression/Monoclonal antibody分类
农业科技引用本文复制引用
孟凡涛,陈芳芳,余为一..鸡新城疫病毒F基因片段的原核表达及其抗F蛋白单克隆抗体的制备[J].安徽农业科学,2012,40(26):12915-12916,2.基金项目
国家自然科学基金项目(30671537). （30671537）
