16S rDNA序列分析法快速鉴定西藏地区传统乳制品中的乳酸菌  

Rapid Identification of Lactic Acid Bacteria from Traditional Dairy Products in Tibet Area by 16S rDNA Sequence Analysis

作　　者：夏雪娟[1] 陈芝兰[2] 陈宗道[1] 阚建全[1] 杨吉霞[1] 

机构地区：[1]西南大学食品科学学院,食品科学与工程实验教学中心,重庆北碚400715 [2]西藏大学农牧学院生物技术中心,西藏林芝860000

出　　处：《食品科学》2013年第14期245-249,共5页Food Science

基　　金：中央高校基本科研业务费专项(XDJK2009C039);国家自然科学基金地方项目(31060020)

摘　　要：对14株乳酸菌进行菌种鉴定。提取基因组DNA,采用属特异性聚合酶链式反应(PCR)、16S rDNA序列同源性分析、种特异性PCR与NEBcutter分析相结合的方法对乳酸菌进行鉴定。结果表明,14株菌中,除123-2和23-3外,其余12株菌均属于乳杆菌属;11-2、13-3、14-3、16-4、23-3、24-3、26-1、122-1、123-4为干酪乳杆菌或副干酪乳杆菌,17-5、20-2、28-1属于植物乳杆菌,123-2为肠膜状明串株菌,125-2为类布氏乳杆菌,除23-3外均与属特异性PCR结果相匹配;11-2、13-3、14-3、16-4、23-3、24-3、26-1、122-1、123-4均为副干酪乳杆菌。NEBcutter分析结果进一步验证了菌株划分的准确性。该方法快速可靠,且误差较小。Objective： Fourteen strains of lactic acid bacteria were identified by a series of taxonomic methods based on 16S rDNA sequence.Methods： Genomic DNAs were extracted from 14 strains.PCR amplification with Lactobacillus genus-specific primers was employed to select Lactobacillus strains.Homology analysis of 16S rDNA sequence,speciesspecific PCR and NEBcutter analysis were combined to indentify 14 strains to species level.Results： Except 123-2 and 23-3 all strains were identified to be Lactobacillus strains by Lactobacillus genus-specific PCR amplification.Through homology analysis of 16S rDNA sequence,strains 11-2,13-3,14-3,16-4,23-3,24-3,26-1,122-1 and 123-4 were assigned to Lactobacillus casei or Lactobacillus paracasei,17-5,20-2 and 28-1 to Lactobacillus plantarum,123-2 to Leuconostoc mesenteroides,125-2 to Lactobacillus parabuchneri.Except for 23-3,the results from homology analysis corresponded well with those obtained from genus-specific PCR.Stains 11-2,13-3,14-3,16-4,23-3,24-3,26-1,122-1 and 123-4 were further confirmed to be Lactobacillus paracasei by species-specific PCR.Finally,these results were verified by NEBcutter.Conclusion： Rapid and accurate identification of natural lactic acid bacteria to species level can be obtained by the combinatorial application of taxonomic methods presented in this study.

关 键 词：乳酸菌 鉴定 属特异性聚合酶链式反应 16SrDNA 种特异性聚合酶链式反应 NEBcutter 

分 类 号：Q939.11[生物学—微生物学]