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首页> 外文期刊>Journal of Clinical Microbiology >Molecular probe for identification of medically important Candida species and Torulopsis glabrata.
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Molecular probe for identification of medically important Candida species and Torulopsis glabrata.

机译:用于鉴定医学上重要的念珠菌和光滑拟杆菌的分子探针。

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摘要

A cloned DNA fragment from Candida albicans containing the gene for the protein actin was used to probe the molecular structure of the actin gene of several medically important yeasts (C. albicans, Candida stellatoidea, Candida tropicalis, Candida pseudotropicalis, Candida krusei, Candida parapsilosis, Candida guilliermondii, and Torulopsis glabrata). Whole-cell DNA from each species was digested with restriction endonucleases, electrophoresed on agarose gels, and transferred to nitrocellulose. Radioactively labeled C. albicans actin gene was hybridized to the DNA fragments on the nitrocellulose. The C. albicans probe produced a strong signal with all of the Candida DNAs tested, indicating considerable conservation of this gene. In addition, the actin genes of all of the species tested were found to have no internal EcoRI or SalI restriction sites. With the exception of C. guilliermondii, all of the species tested had a single internal HindIII recognition site. However, the location of flanking restriction sites was found to be species specific. For all of the enzymes tested, the locations of the flanking restriction sites in C. albicans and C. stellatoidea were identical; all of the other strains yielded fragments clearly distinct from one another. These differences provide a molecular tool for the differentiation of medically important Candida species.
机译:从白色念珠菌克隆的含有DNA肌动蛋白基因的DNA片段被用来探测几种医学上重要的酵母(念珠菌,白色念珠菌,热带念珠菌,拟假丝酵母,克鲁斯念珠菌,副念珠菌,念珠菌念珠菌(Candida guilliermondii)和毛状Torulopsis glabrata)。用限制性核酸内切酶消化每种物种的全细胞DNA,在琼脂糖凝胶上电泳,然后转移到硝酸纤维素膜上。放射性标记的白色念珠菌肌动蛋白基因与硝化纤维素上的DNA片段杂交。白色念珠菌探针对所有测试的念珠菌DNA产生强信号,表明该基因有相当大的保守性。另外,发现所有测试物种的肌动蛋白基因都没有内部EcoRI或SalI限制位点。除吉氏梭菌外,所有测试的物种均具有单个内部HindIII识别位点。然而,发现侧翼限制位点的位置是物种特异性的。对于所有测试的酶,白色念珠菌和恒星念珠菌侧翼限制位点的位置是相同的。所有其他菌株均产生明显彼此不同的片段。这些差异为区分医学上重要的念珠菌提供了分子工具。

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