华北农学报 ›› 2011, Vol. 26 ›› Issue (6): 85-88. doi: 10.7668/hbnxb.2011.06.016

所属专题: 薯类作物

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重组CP多克隆抗体在马铃薯卷叶病毒DAS-ELISA检测中的应用

李楠楠1, 左玉玲1, 隋炯明1, 盖树鹏1, 樊连梅1, 李广存2, 郭宝太1   

  1. 1. 青岛农业大学生命科学学院, 山东省旱作农业技术重点实验室, 山东青岛 266109;
    2. 山东省农业科学院蔬菜研究所, 山东济南 250100
  • 收稿日期:2011-09-14 出版日期:2011-12-28
  • 通讯作者: 郭宝太(1963-),男,山东青州人,教授,博士,主要从事植物分子生物学与基因工程研究。
  • 作者简介:李楠楠(1985-),男,山东淄博人,硕士,主要从事植物基因工程研究。
  • 基金资助:
    山东省自然基金项目(Y2008D43)

Application of Polyclonal Antibody against the Recombinant CP to the DAS-ELISA Detection of Potato Leafroll Virus

LI Nan-nan1, ZUO Yu-ling1, SUI Jiong-ming1, GAI Shu-peng1, FAN Lian-mei1, LI Guang-cun2, GUO Bao-tai1   

  1. 1. College of Life Ssience, Qingdao Agricultural University Key Lab of Dry Farming Fechnology in Shandong Provice, Qingdao 266109, China;
    2. Vegetable Institute, Shandong Academy of Agriculture Science, Jinan 250100, China
  • Received:2011-09-14 Published:2011-12-28

摘要: 先经过饱和硫酸铵沉淀分离,再用Protein G亲和层析柱纯化,从马铃薯卷叶病毒(PLRV)重组CP作抗原制备的抗血清中获得了高纯度多克隆抗体(IgG),并用戊二醛法一步法进行碱性磷酸酶的标记获得了酶标抗体(IgG-AP)。将纯化的抗体与酶标抗体用于感染PLRV病叶的检测,DAS-ELISA反应呈阳性,结果表明,用重组CP制备的多克隆抗体可成功地用于PLRV的DAS-ELISA检测。本研究为PLRV重组CP多克隆抗体的大量制备及ELISA检测奠定了基础。

关键词: 马铃薯卷叶病毒, 重组CP, 多克隆抗体, 酶标抗体, DAS-ELISA

Abstract: Polyclonal antibody(IgG) was isolated from antiserum against the recombinant coat proitein(CP) of PLRV by the treatment of saturated ammmonium sulfate, then crude IgG was purified with protein G affinity chromatography column. High purity IgG was achieved and labelled with alkaline phosphatase by one-step method of glutaraldehyde, a nd enzyme-conjugated IgG(IgG-AP) was obtained. When PLRV-infected potato leaf was detected by DAS-ELISA procedure using the prepared IgG and IgG-AP,positive reaction was observed,and there was no positive reaction in the detection of healthy leaf. The result indicated that the IgG against the recombinant CP and the resulting IgG-AP were successfully used in DAS-ELISA detection of potato leafroll virus.

Key words: Potato leafroll virus(PLRV), Recombinant CP, Polycolonal antibody, Phosphatase-conjugated IgG, DAS-ELISA

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引用本文

李楠楠, 左玉玲, 隋炯明, 盖树鹏, 樊连梅, 李广存, 郭宝太. 重组CP多克隆抗体在马铃薯卷叶病毒DAS-ELISA检测中的应用[J]. 华北农学报, 2011, 26(6): 85-88. doi: 10.7668/hbnxb.2011.06.016.

LI Nan-nan, ZUO Yu-ling, SUI Jiong-ming, GAI Shu-peng, FAN Lian-mei, LI Guang-cun, GUO Bao-tai. Application of Polyclonal Antibody against the Recombinant CP to the DAS-ELISA Detection of Potato Leafroll Virus[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(6): 85-88. doi: 10.7668/hbnxb.2011.06.016.

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