亚热带植物科学 ›› 2009, Vol. 38 ›› Issue (04): 1-4.DOI: 10.3969/j.issn.1009-7791.2009.04.001

• 研究报告 •    下一篇

甘蔗花叶病毒P1基因的原核表达及其抗体制备

郭莺1,2,阮妙鸿2,刘佳2,杨川毓2,张木清2   

  1. 1. 福建省亚热带植物研究所, 福建 厦门 361006;
    2. 农业部甘蔗生理生态与遗传改良重点开放实验室, 福建 福州 350002
  • 收稿日期:2009-08-03 出版日期:2009-11-10 发布日期:2009-11-10
  • 作者简介:郭莺(1981-),女,福建龙岩人,博士,助理研究员,从事植物抗病分子育种研究。
  • 基金资助:

    国家自然科学基金项目(30871514)

Expression and Antibody Preparation of Sugarcane Mosaic Virus P1 Protein

GUO Ying1,2, RUAN Miao-hong2, LIU Jia2, YANG Chuan-yu2, ZHANG Mu-qing2   

  1. 1. Fujian Institute of Subtropical Botany, Xiamen 361006, Fujian China;
    2. Key Lab of Eco-physiology & Genetic Improvement for Sugarcane, Ministry of Agriculture, Fuzhou 350002, Fujian China
  • Received:2009-08-03 Online:2009-11-10 Published:2009-11-10

摘要: 将扩增的甘蔗花叶病毒P1基因克隆到原核表达载体pET-32a上,转化BL21(DE3),获得重组子pET-32a-P1。超声波结果显示,所得的融合蛋白以可溶性的形式存在。SDS-PAGE检测其表达产物与目的蛋白大小一致,割胶免疫注射家兔得到抗体。间接ELISA测定结果表明稀释51200倍仍呈阳性信号,并通过western blot检测,证明抗体特异性良好。

关键词: 甘蔗花叶病毒, P1, 融合蛋白表达, 抗体

Abstract: The target gene was subcloned into the prokaryotic expression vector pET-32a,and transformed into E. coli Bl21(DE3),to obtain recombinant pET-32a-P1.Ultrasound results showed that the fusion protein existed in soluble form.Expression product had the same size with the target protein detected by SDS-PAGE.The antibody against the P1 protein was gained by immunized rabbit with target protein cut from SDS-PAGE.Indirect ELISA measurements proved that the antibody specificity was fine,because of that the dilution of 51200-fold not merely kept positive still,but also passed western blot.

Key words: Sugarcane mosaic virus, P1, fusion protein expression, antibody

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