CESAM & Department of Biology, University of Aveiro, 3810-193 Aveiro, Portugal.
Res Microbiol. 2010 Jan-Feb;161(1):58-66. doi: 10.1016/j.resmic.2009.11.004. Epub 2009 Dec 22.
To investigate the mobile gene pool present in wastewater environments, total community DNA was obtained from two distinct raw effluents: urban and slaughterhouse wastewaters. Bacterial community structure was evaluated by DGGE analysis of 16S rRNA gene fragments. Detection of broad-host-range plasmid sequences and integrase encoding genes was carried out through PCR and Southern hybridization. Gene cassette libraries were obtained using primers targeting consensus sequences that flank gene cassettes. Analysis of DGGE profiles revealed a complex and distinct bacterial community among effluents (similarity<25%). Despite that, All plasmid-specific sequences searched (rep for IncN, trfA for IncP-1 and oriV for IncQ and IncW) and integrase genes were present in both effluents. Gene cassettes recovered from clone libraries showed low homology with genes encoding putative enzymes involved in the metabolism of amino sugars, cell wall synthesis, motility, gene regulation, intercellular signalling and secretion pathways and in the synthesis of cellulose, folic acid and antibiotics. Additionally, in the majority of clones, no identifiable open reading frames homologues were found in the databases. According to our results, wastewater environments promote the development of bacterial communities that support and bring together different types of molecular elements that, in association, play a major role in bacterial adaptation and evolution.
为了研究废水中存在的移动基因库,从两种不同的原废水(城市废水和屠宰废水)中获得了总群落 DNA。通过 16S rRNA 基因片段的 DGGE 分析评估了细菌群落结构。通过 PCR 和 Southern 杂交检测了广谱质粒序列和整合酶编码基因。使用针对基因盒侧翼保守序列的引物获得了基因盒文库。DGGE 图谱分析显示废水中存在复杂而独特的细菌群落(相似性<25%)。尽管如此,在两种废水中均存在搜索到的所有质粒特异性序列(IncN 的 rep、IncP-1 的 trfA、IncQ 和 IncW 的 oriV)和整合酶基因。从克隆文库中回收的基因盒与编码参与氨基酸糖代谢、细胞壁合成、运动、基因调控、细胞间信号传递和分泌途径以及纤维素、叶酸和抗生素合成的假定酶的基因具有低同源性。此外,在大多数克隆中,数据库中未发现可识别的开放阅读框同源物。根据我们的结果,废水环境促进了支持和聚集不同类型分子元素的细菌群落的发展,这些分子元素共同在细菌适应和进化中发挥主要作用。