Environment Education Center, Miyagi University of Education, 149 Aoba, Aramaki, Aoba-ku, Sendai, Miyagi 980-0845, Japan.
Microbes Environ. 2008;23(4):356-9. doi: 10.1264/jsme2.me08532.
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.
通常通过 PCR 扩增和 DNA 测序对纤毛虫进行分子分析，从大量细胞中提取 DNA。因此，纤毛虫的分析仅限于那些可以培养的物种。我们提出了一种单细胞 PCR 程序来克服无法培养物种分析的困难。该程序已针对 6 株 Stichotrichia 菌株和未培养的 Levicoleps biwae 细胞进行了测试，目标是 18S rRNA 基因序列，在进行显微镜观察并获得照片记录后进行。该程序通过将单细胞的形态和基因型联系起来，使对自然环境中无法培养的纤毛虫菌株进行系统分析成为可能。
