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以华南地区典型农村塘坝饮用水为研究对象,采用基于大肠杆菌的两种特异性基因(β-D-葡萄糖苷酶编码基因uidA和膜外周磷通道蛋白编码基因phoE)的群体差异分析(PCR-DGGE),对饮用水中的污染情况进行了溯源研究.结果表明,采用富集培养的混合菌体DNA作为模板扩增大肠杆菌特异性基因更加实用.uidA和phoE的引物均具有较强的特异性.对Genebank中已有的两种特异性基因序列的分析结果表明,phoE的平均变异程度约为uidA的2倍,基于phoE的PCR-DGGE分析技术能够更好地反映样品之间的联系,适合在微生物溯源中应用.研究还表明,塘坝饮用水及其周围环境污染呈现面源特征,养殖废弃物是水体污染的主要来源.
Taking the typical drinking water of ponds in rural areas of southern China as the research object, population-based differential analysis (PCR-RFLP) of two specific genes based on Escherichia coli (the coding gene uidA of β-D-glucosidase and the membrane- DGGE) was used to trace the contamination in drinking water.The results showed that it is more practical to amplify the Escherichia coli-specific genes by using the enriched mixed bacterial DNA as a template.The primers of uidA and phoE have strong specificity The analysis of two specific gene sequences in Genebank showed that the average degree of variation of phoE was about 2 times that of uidA. The PCR-DGGE analysis based on phoE could better reflect the correlation between samples, Suitable for traceability of microorganisms.The study also shows that the drinking water and its surrounding area in Tangba have the characteristics of non-point source pollution, and the culture waste is the main source of water pollution.