Linkage between Light Microscopic Observations and Molecular Analysis by Single-Cell PCR for Ciliates
Linkage between Light Microscopic Observations and Molecular Analysis by Single-Cell PCR for Ciliates
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DOI：
10.1264/jsme2.me08532
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发表时间：
2008-01-01
影响因子：
2.2
通讯作者：
Kasahara, Yasuhro
中科院分区：
文献类型：
--
作者：
Shimano, Satoshi;Sanbe, Mitsuo;Kasahara, Yasuhro
纤毛虫的分子鉴定和分类学研究通常是通过PCR扩增和DNA测序进行的，需要从大量的细胞中提取DNA，因此，纤毛虫的分子鉴定和分类学研究仅限于可培养的种类。我们提出了一个单细胞PCR程序，以克服在分析不可培养的物种的困难。该程序已经测试了6 Stichotrichia菌株和未培养Levicoleps biwae细胞，针对18S rRNA基因序列，进行显微镜观察和获得照片记录后，该程序能够系统分析的自然环境中的不可培养的纤毛虫菌株，通过连接单个细胞的形态和基因型。
The molecular analysis of ciliates for identification and phylogeny is usually conducted through PCR amplification and DNA sequencing, with DNA extracted from a large number of cells, Therefore, the analysis of ciliates is limited to only those species that can be cultured. We propose a single-cell PCR procedure to overcome the difficulty in the analysis of unculturable species. The procedure has been tested on 6 Stichotrichia strains and uncultured Levicoleps biwae cells, targeting 18S rRNA gene sequences, after carrying out microscopic observations and obtaining photographic records, This procedure enables the systematic analysis of unculturable ciliate strains in natural environments by linking the morphology and genotype of a single cell.