中国水稻科学2017，Vol.31Issue(3)：232-237,6.DOI:10.16819/j.1001-7216.2017.6148
水稻齿叶矮缩病毒Pns10蛋白在水稻原生质体内的表达
Expression of Pns10 Protein of Rice ragged stunt virus in Rice Protoplasts
摘要
Abstract
[Objective]Pns10 of Rice ragged stunt virus (RRSV) forms viroplasm-like inclusion bodies which are essential for virus infection to vector insects. However, the expression pattern and function of Pns10 in rice remain unknown.[Method]Polyclonal antibody against Pns10 protein was prepared by immunizing rabbits with a bacterially expressed Pns10; the cellular distribution of Pns10 protein in rice protoplasts was observed with an immunofluorescence microscopy; the accumulation of Pns10 RNA in rice protoplasts infected by RRSV was quantified with real-time quantitative RT-PCR and that of the Pns10 protein was investigated using Western blot.[Result]Pns10 ORF was cloned into the Gateway prokaryotic expression vector Pdest17 and its expression was induced by IPTG. Polyclonal antiserum against the Pns10 was obtained and it can react with the Pns10 protein from the naturally RRSV-infected rice by Western blot detecting. Pns10 formed viroplasm-like inclusion bodies in protoplasts. The Pns10 RNA was first detected at 8 h post RRSV inoculation. The accumulation of Pns10 RNA peaked at 24 h post virus inoculation but began to decline thereafter. The Pns10 protein was first detected at 24 h post virus inoculation. A high level of Pns10 protein was maintained until 60 h post virus inoculation.[Conclusion]Antiserum against RRSV Pns10 protein was obtained. Pns10 was expressed and viroplasm-like inclusion bodies formed in rice protoplasts. In addition, the expression of Pns10 RNA was earlier than that of the Pns10 protein.关键词
水稻齿叶矮缩病毒/水稻原生质体/Pns10抗体/Pns10RNA/Pns10蛋白Key words
Rice ragged stunt virus/rice protoplast/Pns10 antibody/Pns10 RNA/Pns10 protein分类
生物科学引用本文复制引用
张洁,陈晓敏,吴锦鸿,朱重庆,丁新伦,吴祖建..水稻齿叶矮缩病毒Pns10蛋白在水稻原生质体内的表达[J].中国水稻科学,2017,31(3):232-237,6.基金项目
高等学校博士学科点专项科研基金资助项目(20133515120004) （20133515120004）
国家自然科学基金资助项目(31301640) （31301640）
福建教育厅科技项目(JA13103). （JA13103）
