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白色念珠菌中重复序列(RPSs)的染色体定位分析。

Analysis of the chromosomal localization of the repetitive sequences (RPSs) in Candida albicans.

作者信息

Chindamporn A, Nakagawa Y, Homma M, Chibana H, Doi M, Tanaka K

机构信息

Laboratory of Medical Mycology, Nagoya University School of Medicine, Japan.

出版信息

Microbiology (Reading). 1995 Feb;141 ( Pt 2):469-76. doi: 10.1099/13500872-141-2-469.

Abstract

The location and organization of repetitive sequences, members of the RPS family, which are sequences specific to Candida albicans, were determined on each chromosome of C. albicans strain FC18. Using pulsed-field gel electrophoresis, we separated seven fractions from eight chromosomes. Each chromosome was cleaved by BamHI and XhoI to excise the RPSs, which were then detected by hybridization with an RPS probe. All chromosomes except chromosome 4 carried RPSs, and these RPSs were located within a limited region on each chromosome. From the digestion of each chromosome with SfiI and probing with the RPSs, we found that these recognition sites within the RPS region were conserved among all RPS-containing chromosomes. For further characterization of the RPSs, the locations and the boundary regions of the RPSs were examined on chromosome 6 of strain FC18 as a model chromosome. Using the restriction enzymes SfiI, SmaI, XhoI, BamHI, MluI and NruI, we constructed a semi-macro physical map of the RPSs and their boundary regions on this chromosome. We also determined which part of the RPS was adjacent to each boundary by using sub-fragments of RPS as probes. The physical configuration around the RPSs and their boundary regions are presented. The results obtained should be useful for future analysis of the function of these regions.

摘要

确定了白色念珠菌FC18菌株每条染色体上白色念珠菌特异性序列——RPS家族成员重复序列的位置和组织方式。我们使用脉冲场凝胶电泳从八条染色体中分离出七个片段。每条染色体用BamHI和XhoI酶切以切除RPS,然后用RPS探针通过杂交进行检测。除4号染色体外,所有染色体都携带RPS,且这些RPS位于每条染色体的有限区域内。通过用SfiI对每条染色体进行酶切并用RPS进行探针检测,我们发现RPS区域内的这些识别位点在所有含RPS的染色体中都是保守的。为了进一步表征RPS,以FC18菌株的6号染色体作为模型染色体,研究了RPS的位置及其边界区域。我们使用限制性内切酶SfiI、SmaI、XhoI、BamHI、MluI和NruI,构建了该染色体上RPS及其边界区域的半宏观物理图谱。我们还使用RPS的亚片段作为探针,确定了RPS的哪一部分与每个边界相邻。展示了RPS及其边界区域周围的物理构型。所获得的结果应有助于未来对这些区域功能的分析。

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