福建农林大学学报（自然科学版）Issue(1)：42-47,6.DOI:10.13323/j.cnki.j.fafu(nat.sci.).2016.01.008
水稻矮缩病毒非结构蛋白Pns6和外壳蛋白P8多克隆抗体的制备及其应用
Preparation and application of polyclonalantibodies against non-structural protein Pns6 and coat protein P8 of Rice dwarf virus
摘要
Abstract
A full length cDNA of Rice dwarf virus ( RDV) segment S6 encoding movement protein and segment S8 encoding coat pro-tein were cloned from infected rice samples by RT-PCR. Vectors of pDEST17-Pns6 and pDEST17-P8 were expressed via Gateaway system and followed by being transformed into E.coli Rosetta. Then the 59 and 46 ku HIS-tag fusion protein, obtained with the in-duction of IPTG from E.coli Rosetta cells, were used to immunize New Zealand Rabbits as an antigen for polyclonal antibodies pro-duction. Both Western blot and ELISA method proved that both antibodies could detect infected rice plants specifically, indirect ELISA method detection of antiserum titer of Pns6 and P8 were 6 400 times. To summarize, Dot-blot ELISA was established for reli-able, sensitive and specificfor RDV detection. Both antibodies against non-structural protein and coat protein of Rice dwarf virus could be applied to fast virus detection at field-scale.关键词
水稻矮缩病毒/非结构蛋白Pns6/外壳蛋白P8/多克隆抗体/Dot-blot ELISAKey words
Rice dwarf virus/non-structural protein Pns6/coat protein P8/polyclonal antibodies/Dot-blot ELISA分类
农业科技引用本文复制引用
宛柏杰,林文武,吴锦鸿,陈晓敏,吴祖建,张洁..水稻矮缩病毒非结构蛋白Pns6和外壳蛋白P8多克隆抗体的制备及其应用[J].福建农林大学学报（自然科学版）,2016,(1):42-47,6.基金项目
高等学校博士学科点专项科研基金(新教师类)(20133515120004)；国家自然科学基金(31301640)；福建省教育厅科技项目( JA13103) （新教师类）
