Department of Microbiology, Montana State University, Bozeman, Montana 59717.
Appl Environ Microbiol. 1989 Jul;55(7):1818-22. doi: 10.1128/aem.55.7.1818-1822.1989.
Cloning of cDNA obtained from 16S rRNA (16S rcDNA) selectively retrieves species-specific sequence information useful for analyzing the composition and structure of natural microbial communities. With this technique we obtained recombinant 16S rcDNA libraries from Escherichia coli and from a model hot-spring cyanobacterial-mat community. The recombinant plasmids contained exclusively 16S rRNA-derived inserts. This selective approach is independent of biasing culture techniques and eliminates the laborious screening required to locate 16S rRNA gene-bearing recombinants in genomic DNA libraries obtained from natural communities.
从 16S rRNA(16S rDNA)获得的 cDNA 的克隆选择性地回收了用于分析自然微生物群落组成和结构的物种特异性序列信息。使用该技术,我们从大肠杆菌和模型温泉蓝藻垫群落中获得了重组 16S rDNA 文库。重组质粒仅包含 16S rRNA 衍生的插入片段。这种选择性方法独立于有偏向性的培养技术,并且消除了在从自然群落获得的基因组 DNA 文库中定位 16S rRNA 基因携带重组体所需的繁琐筛选。