代表性差异分析比较两株来自不同地区的猪源Lactobacillus菌株  

Comparison of two Lactobacillus strains isolated from piglets in different area by using representational difference analysis

作　　者：苏勇[1] 姚文[1] 朱伟云[1] 

机构地区：[1]南京农业大学消化道微生物研究室,南京210095

出　　处：《微生物学报》2008年第5期577-582,共6页Acta Microbiologica Sinica

基　　金：国家"973项目"(2004CB1175004);国家杰出青年科学基金(30025034)~~

摘　　要：【目的】对分离自猪肠道的乳酸杆菌S1菌株进行鉴定,并比较该菌株与同种的001T菌株的基因差异。【方法】对S1菌株进行16SrRNA基因序列分析和种特异PCR检测,并且对S1菌株和Lactobacillus sobrius 001T进行代表性差异分析(Representational difference analysis,RDA)。【结果】16SrRNA基因序列分析表明,与S1菌株最相似的已知菌为L.sobrius。变性梯度凝胶电泳分析显示,仔猪空、回肠细菌图谱中有一与S1菌株有相同迁移位置的优势条带,克隆、测序鉴定表明,与该条带相匹配的16SrRNA基因克隆(CloneS)的最相似已知菌也为L.sobrius。16SrRNA基因系统进化分析表明,S1菌株与CloneS和L.sobrius16SrRNA基因序列同源性分别为99.8%和99.6%。L.sobrius特异性引物也可以扩增S1株菌的16SrRNA基因的特定片段。因此S1菌株可被确定为L.sobrius。RDA对菌株S1和同种的猪源L.sobrius001T菌株的基因差异进行分析,未发现这两株菌的基因组差异。【结论】猪肠道乳杆酸菌S1菌株属于L.sobrius,其与猪源L.sobrius001T菌株为相似菌株。[Objective] To identify Lactobacillus sp. strain S 1 from the intestine of piglet, and compare the genomic difference between this strain and Lactobacillus sobrius 001^T. [Methods] Analysis of 16S rRNA gene sequence and species-specific polymerase chain reaction assay were used to identify Lactobacillus sp. strain S 1. Representational difference analysis （RDA） was used to compare these two strains. [Results] The 16S rRNA gene sequence analysis of strain S 1 showed that its closest known species in the GenBank database was L. sobrius. Denaturing gradient gel electrophoresis analysis showed that the predominant bands in profiles from bacteria in the jejunum and ileum of piglets had the identical migrating position with strain S 1. Cloning and sequencing of 16S rRNA gene analysis revealed that this band matched clone （Clone S） was also related closely to L. sobrius. Phylogenetic analysis of 16S rRNA gene showed that homology between strain S1 and Clone S was 99.8%, and that between strain S1 and L. sobrius was 99.6%. Both strains could be detected in 1.2% agarose gel by L. sobrius-specific PCR assay. Recently, RDA has been adapted to study the genomic diversity of bacterial strains. This analysis showed that there was no genomic difference between the two strains. [Conclusion] Piglet-derived strain S1 belonged to L. sobrius. Piglet-derived L. sobrius 001^T and L. sobrius S1 were the similar strain.

关 键 词：Lacwbacillus sobrius 代表性差异分析 猪肠道乳酸杆菌 

分 类 号：S852.6[农业科学—基础兽医学]