National Institute of Livestock and Grassland Science, Tsukuba, Ibaraki, Japan.
FEMS Microbiol Lett. 2011 Mar;316(1):70-6. doi: 10.1111/j.1574-6968.2010.02193.x. Epub 2011 Jan 24.
Lactobacillus paraplantarum is a species phenotypically close to Lactobacillus plantarum. Several PCR methods were evaluated to discriminate L. paraplantarum strains and among them, a PCR using an enterobacterial repetitive intergenic consensus (ERIC) sequence differentiated L. paraplantarum from other Lactobacillus species. In addition, a combination of ERIC and random amplified polymorphic DNA (RAPD) analysis distinguished among seven strains of L. paraplantarum tested. ERIC-PCR profiles showed several strain-specific DNA fragments in L. paraplantarum, among them, a 2.2-kb ERIC marker, termed LpF1, found to be specific to strain FBA1, which improved the skin integrity in an animal model. The LpF1 encodes three proteins similar to Lactobacillus fermentum AroA, TyrA, and AroK, which are involved in the shikimate pathway. A primer pair specific to FBA1 based on the internal sequence of LpF1 amplified a 950-bp FBA1-specific fragment LpF2. Southern blot analysis of Dra I-digested genomic DNA of L. paraplantarum strains using LpF2 as a probe showed that LpF2 is distinctive of strain FBA1 among 16 L. paraplantarum strains. Because both ERIC- and RAPD-PCR are fast and technically simple methods, they are useful for the rapid discrimination of L. paraplantarum strains and for the development of new strain-specific DNA markers for identifying industrially important strains.
副干酪乳杆菌在表型上与植物乳杆菌非常接近。我们评估了几种 PCR 方法来区分副干酪乳杆菌菌株，其中使用肠细菌重复基因间一致（ERIC）序列的 PCR 可将副干酪乳杆菌与其他乳杆菌物种区分开来。此外，ERIC 和随机扩增多态性 DNA（RAPD）分析的组合可区分测试的 7 株副干酪乳杆菌。ERIC-PCR 图谱显示副干酪乳杆菌中有几个菌株特异性 DNA 片段，其中一个 2.2-kb 的 ERIC 标记，称为 LpF1，被发现是 FBA1 菌株特异性的，可改善动物模型中的皮肤完整性。LpF1 编码三个与乳酸乳杆菌 AroA、TyrA 和 AroK 相似的蛋白，这些蛋白参与莽草酸途径。基于 LpF1 内部序列设计的针对 FBA1 的引物对可扩增出 950-bp 的 FBA1 特异性片段 LpF2。使用 LpF2 作为探针对 Dra I 消化的副干酪乳杆菌菌株基因组 DNA 进行 Southern blot 分析表明，LpF2 是 16 株副干酪乳杆菌菌株中 FBA1 菌株的独特标记。由于 ERIC 和 RAPD-PCR 都是快速且技术简单的方法，因此它们可用于快速区分副干酪乳杆菌菌株，并开发用于鉴定工业重要菌株的新菌株特异性 DNA 标记。
