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首页> 外文期刊>Bioscience, Biotechnology, and Biochemistry >Screening for a gene deletion mutant whose temperature sensitivity is suppressed by FK506 in budding yeast and its application for a positive screening for drugs inhibiting calcineurin
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Screening for a gene deletion mutant whose temperature sensitivity is suppressed by FK506 in budding yeast and its application for a positive screening for drugs inhibiting calcineurin

机译：筛选用于培养酵母的FK506在萌芽酵母中抑制温度敏感性的基因缺失突变体及其在抑制钙素素的药物阳性筛选中的应用

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摘要

Calcineurin, which is a Ca2+/calmodulin-dependent protein phosphatase, is a key mediator in calcium signaling in diverse biological processes and of clinical importance as the target of the immunosuppressant FK506. To identify a mutant(s) in which calcineurin is activated, inhibiting cellular growth as a result, we screened for a mutant(s) whose temperature sensitivity would be suppressed by FK506 from the budding yeast non-essential gene deletion library. We found that the temperature sensitivity of cells in which the conserved Verprolin VRP1 gene had been deleted, which gene is required for actin organization and endocytosis, was suppressed by either FK506 or by cnb1 deletion. Indeed, the calcineurin activity increased significantly in the increment vrp1 cells. Finally, we demonstrated that the increment vrp1 strain to be useful as an indicator in a positive screening for bioactive compounds inhibiting calcineurin.
机译：作为Ca2 + /钙调蛋白依赖性蛋白质磷酸酶的煅烧肽是在不同的生物过程中钙信号传导的关键介质，并且作为免疫抑制剂FK506的靶标。 为了鉴定其中激活钙癌素的突变体，抑制细胞生长，我们筛选突变体，其温度敏感性来自萌芽酵母非必需基因缺失文库的FK506将被FK506抑制。 我们发现，通过FK506或CNB1缺失抑制了脱蛋白组织和内吞作用所需的细胞的温度敏感性。 实际上，增量VRP1细胞的钙素活性显着增加。 最后，我们证明，增量VRP1应变作为抑制钙素蛋白的生物活性化合物的正筛选中的指标。

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