ERIC-PCR在人工混合菌体系中的检出灵敏度  

The Sensitivity of ERIC-PCR in the Mixed Bacteria System

作　　者：李艳琴[1] 孙永艳[1] 崔丽方[1] 申泉[1] 

机构地区：[1]山西大学生物技术研究所化学生物学与分子工程教育部重点实验室,太原030006

出　　处：《微生物学通报》2004年第4期61-64,共4页Microbiology China

基　　金：山西省自然科学基金资助项目 (No 2 0 0 2 1 0 80 )

摘　　要：提取大肠杆菌DH5α和阴沟肠杆菌 (E 2 6R)的基因组DNA ,进行ERIC PCR ,可以分别得到稳定而独特的DNA指纹图谱 ;模板量的变动范围从 1 0～ 1 0 0ng都不会影响其图谱的稳定性 ;在图谱中DH5α和E 2 6R各自均有一条含量最大的特征带 ,其不易受实验环境和实验条件的干扰而发生变化。把DH5α和E 2 6R按比例混合 ,提取混合菌基因组DNA ,进行ERIC PCR ,结果表明 :混合菌的DNA指纹图谱为各纯菌图谱的叠加 ,亦具有稳定性 ;通过对凝胶电泳图谱中特征带的分析 ,可以看出 :当DH5α的菌含量达到总菌量的 0 5 %时 ,可被ERIC PCR方法清楚地检测出来。为ERIC PCR用于土壤微生物和环境微生物的研究 。Genomic DNA of the two bacterial strains including Escherichia coli DH5α and Enterobacter cloacae E 26R were amplified by PCR with specific primers of ERIC sequence Each strain showed the stable and unique DNA fingerprint when PCR products were analyzed in agarose gel electrophoresis The stability of the genomic DNA fingerprint wasn't influenced by the templet from 10ng to 100ng In the fingerprints there were characteristic bands The unique band was influenced difficultly due to the change of the environment and condition of experiment DH5α and E 26R were mixed proportionately, and examined by ERIC PCR The result showed that the DNA fingerprints of the mixture were the superposition of each pure bacterium's Analysis of the main bands showed that DH5α can be examined by ERIC PCR when its concentration gets to 0 5% of the total mixed bacteria The study provides the reference for the research of soil microbe and environmental microbe, and especially can be used for quick identification and examination of the sundry bacterial contamination in the fermentation industry

关 键 词：ERIC-PCR 指纹图谱 混合菌 检出灵敏度 

分 类 号：Q93[生物学—微生物学]