Nübel U, Garcia-Pichel F, Muyzer G
Max Planck Institute for Marine Microbiology, Bremen, Germany.
Appl Environ Microbiol. 1997 Aug;63(8):3327-32. doi: 10.1128/aem.63.8.3327-3332.1997.
We developed and tested a set of oligonucleotide primers for the specific amplification of 16S rRNA gene segments from cyanobacteria and plastids by PCR. PCR products were recovered from all cultures of cyanobacteria and diatoms that were checked but not from other bacteria and archaea. Gene segments selectively retrieved from cyanobacteria and diatoms in unialgal but nonaxenic cultures and from cyanobionts in lichens could be directly sequenced. In the context of growing sequence databases, this procedure allows rapid and phylogenetically meaningful identification without pure cultures or molecular cloning. We demonstrate the use of this specific PCR in combination with denaturing gradient gel electrophoresis to probe the diversity of oxygenic phototrophic microorganisms in cultures, lichens, and complex microbial communities.
我们设计并测试了一组寡核苷酸引物,用于通过聚合酶链反应(PCR)从蓝细菌和质体中特异性扩增16S rRNA基因片段。从所有检测的蓝细菌和硅藻培养物中回收了PCR产物,但未从其他细菌和古细菌中回收。从单种但不纯培养的蓝细菌和硅藻以及地衣中的蓝共生体中选择性获取的基因片段可以直接测序。在不断增长的序列数据库背景下,该方法无需纯培养或分子克隆即可快速且在系统发育上进行有意义的鉴定。我们展示了这种特异性PCR与变性梯度凝胶电泳相结合的用途,以探测培养物、地衣和复杂微生物群落中光合放氧微生物的多样性。