Akimitsu K, Hart L P, Walton J D, Hollingsworth R
Department of Botany and Plant Pathology, Michigan State University, East Lansing, Michigan 48824.
Plant Physiol. 1992 Jan;98(1):121-6. doi: 10.1104/pp.98.1.121.
Polyclonal antibodies against victorin, the host-specific toxin produced by Cochliobolus victoriae, were raised in rabbits immunized with a victorin-bovine serum albumin conjugate. The antibodies were purified from serum by protein A column chromatography and characterized by indirect and direct enzymelinked immunosorbent assays (ELISA). The concentration of victorin that inhibited anti-victorin antibody binding by 50% was 10 nanograms per milliliter in an indirect ELISA. The lowest concentration of victorin detectable was 10 picograms per milliliter. In a direct ELISA, 25 nanograms per milliliter of victorin inhibited binding of victorin-horseradish peroxidase conjugate by 50%. In vivo and in vitro covalent binding of victorin to proteins in susceptible and resistant oat (Avena sativa) tissue was examined by western blotting assays using anti-victorin antibody and a second antibody conjugated with (125)I or alkaline phosphatase. In vivo binding of victorin to proteins of 100 and 45 kilodaltons was observed in both susceptible and resistant cultivars of oats. Victorin also bound in vitro to proteins of 100, 65, and 45 kilodaltons in both susceptible and resistant oats. The data indicate that victorin binds covalently to the same sites in susceptible and resistant genotypes of oats.
用维多利亚长蠕孢菌产生的宿主特异性毒素维克托菌素（victorin）与牛血清白蛋白的偶联物免疫家兔，制备了抗维克托菌素的多克隆抗体。通过蛋白A柱色谱从血清中纯化抗体，并通过间接和直接酶联免疫吸附测定（ELISA）进行表征。在间接ELISA中，抑制抗维克托菌素抗体结合50%的维克托菌素浓度为每毫升10纳克。可检测到的维克托菌素最低浓度为每毫升10皮克。在直接ELISA中，每毫升25纳克的维克托菌素可抑制维克托菌素-辣根过氧化物酶偶联物结合50%。使用抗维克托菌素抗体和与（125）I或碱性磷酸酶偶联的二抗，通过蛋白质印迹分析检测维克托菌素在敏感和抗性燕麦（ Avena sativa）组织中与蛋白质的体内和体外共价结合。在燕麦的敏感和抗性品种中均观察到维克托菌素在体内与100和45千道尔顿的蛋白质结合。维克托菌素在体外也与敏感和抗性燕麦中100、65和45千道尔顿的蛋白质结合。数据表明，维克托菌素在燕麦的敏感和抗性基因型中与相同位点共价结合。
