Molecular probe for identification of medically important Candida species and Torulopsis glabrata.
Molecular probe for identification of medically important Candida species and Torulopsis glabrata.
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用于鉴定医学上重要的念珠菌属和光滑球拟酵母的分子探针。
DOI：
10.1128/jcm.25.3.563-566.1987
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发表时间：
1987
影响因子：
9.4
通讯作者：
Riggsby,WS
中科院分区：
文献类型：
--
作者：
Mason,MM;Lasker,BA;Riggsby,WS
利用从白色念珠菌中克隆的含有肌动蛋白基因的DNA片段，研究了几种医学上重要的酵母(白色念珠菌、星状念珠菌、热带念珠菌、拟热带念珠菌、克鲁斯假丝酵母、近缘假丝酵母菌、假丝酵母菌和光滑念珠菌)的肌动蛋白基因的分子结构。每个物种的全细胞DNA用限制性内切酶消化，琼脂糖凝胶电泳，然后转移到硝酸纤维素上。将放射性标记的白念珠菌肌动蛋白基因与硝酸纤维素膜上的DNA片段杂交。白色念珠菌的探针产生了一个强烈的信号，所有的念珠菌DNA都进行了测试，这表明这个基因相当保守。此外，所有受试物种的肌动蛋白基因都没有EcoRI或SalI限制性内切酶切点。除了Guilliermondii外，所有被测试的物种都有一个单一的内部HindIII识别位点。然而，侧翼限制位点的位置被发现是物种特有的。对于所有测试的酶，白色念珠菌和星形念珠菌的侧翼限制性内切酶位点的位置是相同的；所有其他菌株产生的片段彼此明显不同。这些差异为区分医学上重要的念珠菌物种提供了分子工具。
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.