应用PCR-DGGE研究饮用水中微生物的多样性  

Application of PCR-DGGE in the Research of Microbial Diversity in Drinking Water

作　　者：吴卿[1] 赵新华[1] 

机构地区：[1]天津大学环境科学与工程学院,天津300072

出　　处：《南开大学学报（自然科学版）》2007年第3期92-96,共5页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：国家"十五"重大科技专项(2002AA601120);国家自然科学基金(50478086)

摘　　要：变性梯度凝胶电泳技术(DGGE)在微生物生态学领域有着广泛的应用.应用冻融法直接提取不同饮用水水样微生物基因组 DNA,选择细菌通用性引物 EUBf_(933)GC 和 EUBr_(1387),对包括细菌 V6、7及8区的16SrRNA 基因进行扩增,经变性梯度凝胶电泳(DGGE)进行分离后,得到不同数目且分离效果较好的电泳条带.结果表明,DGGE 能够对饮用水样品中的不同微生物的16S rRNA 基因的 DNA 扩增片段进行分离,为进一步对这些 DNA 片段回收和测序奠定了基础.不同取样点饮用水样品中虽然存在特异菌,但各取样点水样中的优势菌相同.从南北方两个城市水样微生物组成来看,优势菌群基本一致,但南方某市的饮用水微生物学相关指标的测定结果明显好于北方某市,优势菌的数量和微生物种类明显少于北方某市.Denaturing gradient gel electrophoresis （DGGE） has been widely used in the field of microbial ecology in recent years. DNA of bacteria in the drinking water was directly extracted without culture. 16S ribosomal DNA fragments, including V6,-7, and -8 regions, were amplified with universal primers （EUBf933GC and EUBr1387） and analyzed by DGGE. The spectrum of DGGE indicated that amplification products could be separated well. The results showed that DGGE could be used in the separation of different microbial 16S rRNA gene that was extracted from drinking water. Though there were special bacteria in different water samples, the predominance bacteria were essentially the same. The predominance bacteria between the northern city and the southern city were the same. The bacteriological water quality of the southern city is better than that of the northern city, and the quantity species of bacteria are fewer than that of the northern city.

关 键 词：饮用水 变性梯度凝胶电泳(DGGE) 16S RRNA 微生物多样性 

分 类 号：X172[环境科学与工程—环境科学]