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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >Production of monoclonal antibodies against peripheral-vesicle proteins in zoospores of Phytophthora nicotianae
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Production of monoclonal antibodies against peripheral-vesicle proteins in zoospores of Phytophthora nicotianae

机译：烟草疫霉菌游动孢子中针对外周囊泡蛋白的单克隆抗体的产生

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摘要

A coimmunisation protocol using microsomal fractions from Phytophthora nicotianae cells has enhanced the production of monoclonal antibodies directed towards proteins produced during asexual sporulation. Over 40% of the antibodies targeted three categories of zoospore peripheral vesicles. Five antibodies label the contents of dorsal vesicles, with three of these reacting with two P nicotianae polypeptides with a relative molecular mass of approximately 100 kDa. Two antibodies label the contents of large peripheral vesicles and react with two very high-molecular-weight polypeptides in extracts of P nicotianae cells. These antibodies cross-react with the contents of large peripheral vesicles in P. cinnamomi zoospores. Ten antibodies label the contents of P nicotianae zoospore ventral vesicles and react with a single polypeptide with a relative molecular mass of 230 kDa. A number of these antibodies against the contents of ventral vesicles in P nicotianae zoospores cross-react with ventral-vesicle proteins in P cinnamomi cells in immunofluorescence and immunoblot assays. The study illustrates the value of the coimmunisation protocol and has produced antibodies that could be instrumental in the cloning, of genes encoding peripheral-vesicle proteins.
机译：使用来自烟草疫霉菌的微粒体级分的共免疫方案已提高了针对无性孢子形成过程中产生的蛋白质的单克隆抗体的产生。超过40％的抗体靶向三类游动孢子外围囊泡。五个抗体标记了背囊的内容，其中三个与相对分子质量约为100 kDa的两个P烟草多肽反应。两种抗体标记了大的外周囊泡的内容，并与烟草单核细胞提取物中的两种非常高分子量的多肽发生反应。这些抗体与肉桂假单胞菌游动孢子中大外围囊泡的内容物发生交叉反应。十种抗体标记了烟孢子虫游动子腹小泡的内容，并与相对分子质量为230 kDa的单个多肽发生反应。在免疫荧光法和免疫印迹法中，许多针对烟草斜纹夜蛾游动孢子中腹泡囊泡含量的抗体与P肉桂细胞中的腹泡囊泡蛋白发生交叉反应。这项研究说明了共免疫方案的价值，并产生了可能在克隆中编码外围囊泡蛋白基因的抗体。

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