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利用M13介导的RAPD-PCR指纹数据库快速鉴定乳源乳酸菌

Rapid identification of dairy lactic acid bacteria by M13-generated, RAPD-PCR fingerprint databases.

作者信息

Rossetti Lia, Giraffa Giorgio

机构信息

Istituto Sperimentale Lattiero Caseario, Via Lombardo 11, 26900 Lodi, Italy.

出版信息

J Microbiol Methods. 2005 Nov;63(2):135-44. doi: 10.1016/j.mimet.2005.03.001.

Abstract

About a thousand lactic acid bacteria (LAB) isolated from dairy products, especially cheeses, were identified and typed by species-specific PCR and RAPD-PCR, respectively. RAPD-PCR profiles, which were obtained by using the M13 sequence as a primer, allowed us to implement a large database of different fingerprints, which were analysed by BioNumerics software. Cluster analysis of the combined RAPD-PCR fingerprinting profiles enabled us to implement a library, which is a collection of library units, which in turn is a selection of representative database entries. A library unit, in this case, can be considered to be a definable taxon. The strains belonged to 11 main RAPD-PCR fingerprinting library units identified as Lactobacillus casei/paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus helveticus, Lactobacillus delbrueckii, Lactobacillus fermentum, Lactobacillus brevis, Enterococcus faecium, Enterococcus faecalis, Streptococcus thermophilus and Lactococcus lactis. The possibility to routinely identify newly typed, bacterial isolates by consulting the library of the software was valued. The proposed method could be suggested to refine previous strain identifications, eliminate redundancy and dispose of a technologically useful LAB strain collection. The same approach could also be applied to identify LAB strains isolated from other food ecosystems.

摘要

通过物种特异性聚合酶链反应(PCR)和随机扩增多态性DNA聚合酶链反应(RAPD-PCR)分别对从乳制品尤其是奶酪中分离出的约1000株乳酸菌(LAB)进行了鉴定和分型。使用M13序列作为引物获得的RAPD-PCR图谱使我们能够建立一个包含不同指纹的大型数据库,并通过BioNumerics软件进行分析。对合并后的RAPD-PCR指纹图谱进行聚类分析,使我们能够建立一个文库,该文库是文库单元的集合,而文库单元又是代表性数据库条目的选择。在这种情况下,一个文库单元可被视为一个可定义的分类单元。这些菌株属于11个主要的RAPD-PCR指纹文库单元,分别被鉴定为干酪乳杆菌/副干酪乳杆菌、植物乳杆菌、鼠李糖乳杆菌、瑞士乳杆菌、德氏乳杆菌、发酵乳杆菌、短乳杆菌、粪肠球菌、屎肠球菌、嗜热链球菌和乳酸乳球菌。通过查阅软件文库来常规鉴定新分型的细菌分离株的可能性受到重视。该方法可用于完善先前的菌株鉴定,消除冗余,并处理一个技术上有用的LAB菌株集合。同样的方法也可用于鉴定从其他食物生态系统中分离出的LAB菌株。

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