Mason M M, Lasker B A, Riggsby W S
J Clin Microbiol. 1987 Mar;25(3):563-6. doi: 10.1128/jcm.25.3.563-566.1987.
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.
一个来自白色念珠菌的含有肌动蛋白基因的克隆DNA片段被用于探测几种医学上重要酵母（白色念珠菌、星状念珠菌、热带念珠菌、假热带念珠菌、克鲁斯念珠菌、近平滑念珠菌、季也蒙念珠菌和光滑球拟酵母）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，在琼脂糖凝胶上进行电泳，然后转移到硝酸纤维素膜上。用放射性标记的白色念珠菌肌动蛋白基因与硝酸纤维素膜上的DNA片段杂交。白色念珠菌探针与所有测试的念珠菌DNA都产生了强烈信号，表明该基因具有相当高的保守性。此外，所有测试物种的肌动蛋白基因都没有内部EcoRI或SalI限制性酶切位点。除季也蒙念珠菌外，所有测试物种都有一个单一的内部HindIII识别位点。然而，侧翼限制性酶切位点的位置是物种特异性的。对于所有测试的酶，白色念珠菌和星状念珠菌中侧翼限制性酶切位点的位置是相同的；所有其他菌株产生的片段明显彼此不同。这些差异为区分医学上重要的念珠菌物种提供了一种分子工具。
