免疫化学定量分析无黄化衰老突变体羊茅叶片中的细胞色素 f。

Immunochemical quantification of cytochrome f in leaves of a non-yellowing senescence mutant of Festuca pratensis.

机构信息

Welsh Plant Breeding Station, Cell Biology Department, AFRC Institute for Grassland and Animal Production, Plas Gogerddan, SY23 3EB, Aberystwyth, Dyfed, UK.

出版信息

Photosynth Res. 1990 Apr;24(1):99-108. doi: 10.1007/BF00032649.

DOI:10.1007/BF00032649

PMID:24419770

Abstract

A non-competitive enzyme-linked immunosorbent assay (ELISA) which enables detection of as little as 0.1 ng cytochrome f in leaf extracts has been developed. No evidence for specific or non-specific interference by proteins other than cytochrome f was found. The assay was applied to a comparative study of age-related changes in the cytochrome f content of leaves of Festuca pratensis Huds. cv. Rossa, and a non-yellowing mutant genotype (Bf993) having a lesion in the mechanism responsible for thylakoid membrane disassembly. Cytochrome f in senescent leaves of the latter genotype was found to be present at significantly higher levels than in the wild-type, implying an inability on the part of the mutant to degrade this protein. The results obtained by ELISA were confirmed by antibody probing of Western blots.

摘要

已开发出一种非竞争酶联免疫吸附测定（ELISA），可检测低至 0.1ng 的叶提取物中的细胞色素 f。未发现细胞色素 f 以外的蛋白质存在特异性或非特异性干扰。该测定法应用于对草地早熟禾叶片中细胞色素 f 含量随年龄变化的比较研究，以及一种在负责类囊体膜解体的机制中存在缺陷的非黄化突变基因型（Bf993）。发现后者基因型衰老叶片中的细胞色素 f 含量明显高于野生型，这意味着突变体无法降解这种蛋白质。ELISA 获得的结果通过 Western blot 抗体探测得到了证实。