Greene E Anne, Voordouw Gerrit
Department of Biological Sciences, University of Calgary, Calgary, Alberta, Canada T2N 1N4.
J Microbiol Methods. 2003 May;53(2):211-9. doi: 10.1016/s0167-7012(03)00024-1.
Development of fast and accurate methods for monitoring environmental microbial diversity is one of the great challenges in microbiology today. Oligonucleotide probes based on 16S rRNA sequences are widely used to identify bacteria in the environment. However, the successful development of a chip of immobilized 16S rRNA probes for identification of large numbers of species in a single hybridization step has not yet been reported. In reverse sample genome probing (RSGP), labelled total community DNA is hybridized to arrays in which genomes of cultured microorganisms are spotted on a solid support in denatured form. This method has provided useful information on changes in composition of the cultured component of microbial communities in oil fields, the soil rhizhosphere, hydrocarbon-contaminated soils and acid mine drainage sites. Applications and limitations of the method, as well as the prospects of extending RSGP to cover also the as yet uncultured component of microbial communities, are evaluated.
开发快速准确的环境微生物多样性监测方法是当今微生物学面临的重大挑战之一。基于16S rRNA序列的寡核苷酸探针被广泛用于鉴定环境中的细菌。然而，尚未有关于在单个杂交步骤中用于鉴定大量物种的固定化16S rRNA探针芯片成功开发的报道。在反向样品基因组探测（RSGP）中，标记的总群落DNA与阵列杂交，在该阵列中，培养微生物的基因组以变性形式点样在固体支持物上。该方法为油田、土壤根际、烃污染土壤和酸性矿山排水点微生物群落培养组分的组成变化提供了有用信息。对该方法的应用和局限性，以及将RSGP扩展到也涵盖微生物群落中尚未培养的组分的前景进行了评估。
