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专一识别脱落酸甲酯的单克隆抗体的制备与应用

周燮, 郑志富, 陈溥言, 章迪

南京农业大学农学系.江苏,南京 210095；南京农业大学动物医学系.江苏,南京 210095；南京市农业科学研究所.江苏,南京 210046

摘要

专一识别2-顺(S)ABA甲酯的单克隆抗体来源于以ABA分子中的1-COOH为偶联位点合成的免疫原。它与游离态ABA和结合态ABA葡萄糖酯的交叉反应仅分别为1%与3.5%,而与ABA类似物,如2-顺-黄质醛、紫黄质以及ABA的2-反式异构体和(R)-对映体则无交叉反应。利用该抗体建立的高度灵敏和精确的ABAme酶联免疫测定法,其检测线性范围为0.048-1.52pmol。通过ABAme ELISA和GA_(1+3)ELISA分析可知羊蹄叶片衰老与内源GA_(1+3)/ABA比值的下降有关。

关键词： 脱落酸; 赤霉素; 单克隆抗体; 酶联免疫测定法; 羊蹄; 衰老

Preparation and application of monoclonal antibodies specific for abscisic acid methyl ester

Zhou Xie, Zheng Zhifu, Chen Puyan, Zhang Di

Departments of Agronomy,Nanjing Agricultural University.Nanjing 210095,Jiangsu；China；Nanjing Agricultural Institute.Nanjing 210046,Jiangsu

Abstract

A monoclonal antibody 5 E 11-D7-51, IgG_(1), which specifically recognizes abscisic acid methyl ester (ABAme) is described. This antibody was derived from an immunogen in which the carrier proteins were linked to ABA at 1-COOH. It showed very little cross-reactivity with ABA (1%) or ABA glucosyl ester (3.5%). Other compounds structurally similar to ABA, such as 2-cis-xanthoxin, violaxanthin and both the 2, trans-isomer and the (R)-enantiomer of ABA, were not found to exhibit any cross-reactivity (Table 1). Based on the use of this antibody, an enzyme-linked immunosorbent assay (ELISA) with high sensitivity and precision was developed, which displayed a linear range of 0.048-1.52 pmol of ABAme. Using this ELISE, as well as an established ELISA for gibberellins (GA_(1+3)), the levels of ABA and GA_(1+3) in senescing leaves of Rumex japonicus were examined. The results showed that the decrease in GA_(1+3)/ABA ratio was connected with the senescence of leaves of Rumex japonicus.

Key words: Abscisic acid (ABA);Gibberellin (GA);Monoclonal antibody (MAb);Enzyme-linked immunosorbent assay (ELISA);Rumex japonicus;Senescence

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引用本文：

周燮, 郑志富, 陈溥言, 章迪. 专一识别脱落酸甲酯的单克隆抗体的制备与应用[J]. 生理学报 1996; 48 (3): .

Zhou Xie, Zheng Zhifu, Chen Puyan, Zhang Di. Preparation and application of monoclonal antibodies specific for abscisic acid methyl ester. Acta Physiol Sin 1996; 48 (3): (in Chinese with English abstract).