16S rDNA-RFLP技术鉴定西藏地区乳制品中的乳杆菌

将16S rDNA PCR技术和RFLP技术相结合,对分离自乳制品中的乳杆菌进行分类和鉴定.从我国西藏地区传统发酵乳中分离出51株乳杆菌,采用通用引物扩增16S rDNA,利用限制性内切酶AluI,HaeIII和HinfI将16S rDNA扩增产物进行酶切后,经聚丙烯酰胺凝胶电泳图谱将菌种鉴定到种的水平.根据16S rDNA-RFLP的结果从中选出8株有代表性的菌株进行生理生化实验和16S rDNA序列的测定,实验结果与RFLP鉴定结果相吻合,表明此方法是一种快速、准确的可用于大量乳杆菌分类和鉴定的方法.

Identification of Lactobacillus Isolated from Home-Made Fermented Milk in Tibet by 16S rDNA-RFLP

In this study, Lactobacilli were rapidly classified and identified by the combinational method of 16S rDNA PCR and Restriction Fragment Length Polymorphism (RFLP). Fifty one strains of Lactobacillus were isolated from traditional homemade fermented milk in Tibet, China. The 16S rDNA of all isolates were amplified by universal primers, and amplification productions were digested using a set of restriction enzymes, AluI, HaeIII and HinfI. Polyacrylamide gel electrophoregram analysis indicated that the lactobacilli could be clearly identified at the species level. Further studies on 16S rDNA sequences, physiological and biochemical test of 8 different strains showed that 16S rDNA PCR-RFLP analysis is rapid and easy way to identify large scale of Lactobacillus.