摘要
以碱加热裂解法从山西农业大学污水直接和间接排放处理池自然水样中分离细菌总DNA ,以细菌 16SrRNA基因特异性引物扩增 16SrRNA基因片段。采用T -A克隆系统克隆分子量为 1339bp的PCR产物 ,构建了rDNA亚克隆文库。用三种识别六碱基的限制性内切酶两两组合消化重组质粒。共分析了两个处理池自然水样各 30个rDNA重组质粒 ,分别发现 4种和 8种细菌rDNA基因型。各基因型频率的分布变化比较大 ,最高和最低的频率分别为 0 2 5和 0 0 1。部分基因型频率间差异显著 ,遗传多样性指数分别为 0 4和 0 9。
Total DNA from natural water samples of two sewage farms where sewage was let using direct and indirect ways, were isolated with alkaline lysis procedure under 100℃. 16S rDNA fragments are amplified using bacteria specific primers and amplified products are cloned with the T-A Cloning System. 60 Recombinants were digested with 3 restriction enzymes. EcoRI and HindIII is our choice basing on the frequencies and distribution of DNA bands. 4 and 8 RFLP types were detected sampled from different natural samples respectively among 60 recombinants containing full length inserts. The frequencies of rDNA genotypes distributed in a narrow range. Some of differences between two extreme frequencies are dominant, the highest and lowest frequencies are 0 25 and 0 01 respectively. Genetic diversity index is 1 029 for this sample. We predicted that the index of molecular information is positively related to the diversity of bacterial community in sewage poll environment.
基金
山西省青年基金资助项目 (991 0 38)
