Purification and immunological characterization of toxic components from cultures of Pyrenophora tritici-repentis.
Purification and immunological characterization of toxic components from cultures of Pyrenophora tritici-repentis.
复制标题
小麦核腔菌培养物中有毒成分的纯化和免疫学表征。
DOI：
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发表时间：
1995
影响因子：
3.5
通讯作者：
L. Ciuffetti
中科院分区：
文献类型：
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作者：
R. P. Tuori;T. Wolpert;L. Ciuffetti
为了便于小麦-小核腔菌互作致病性的遗传分析，从该真菌培养物中纯化了一种寄主选择性毒性蛋白，命名为ToxA。ToxA被证明是一种13.2 kDa的热稳定蛋白，在敏感的小麦品种中诱导可见坏死，平均最低浓度为60 nM。针对ToxA的多克隆抗体通过Western分析和间接免疫沉淀显示对该蛋白质具有特异性。免疫沉淀蛋白和从聚丙烯酰胺凝胶上洗脱的ToxA蛋白的生物测定表明，ToxA蛋白是毒性剂。在P. tritici-repentis的培养物中也检测到其他在色谱和免疫学上与ToxA不同的不太丰富的坏死诱导组分。这些成分被发现在阳离子和阴离子蛋白组分，像毒素A，诱导特定的坏死。
To facilitate the genetic analysis of pathogenicity in the wheat-Pyrenophora tritici-repentis interaction, a host-selective toxic protein, designated ToxA, was purified from culture filtrates of this fungus. ToxA was shown to be a 13.2-kDa heat-stable protein which induced visible necrosis in sensitive wheat cultivars at an average minimum concentration of 60 nM. Polyclonal antibodies raised against ToxA were shown by Western analysis and indirect immunoprecipitation to be specific for this protein. Bioassays of immunoprecipitated protein and ToxA protein eluted from polyacrylamide gels indicated that ToxA protein is the toxic agent. Other less abundant necrosis-inducing components that are chromatographically and immunologically distinct from ToxA were also detected in culture filtrates of P. tritici-repentis. These components were found in cationic and anionic protein fractions and, like ToxA, induced cultivar-specific necrosis.