16S rDNA-RFLP技术鉴定西藏地区乳制品中的乳杆菌  

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

作　　者：于洁[1] 孙志宏[1] 张家超[1] 艾日登才次克[1] 张彦斌[1] 杨梅[1] 孙天松[1] 张和平[1] 

机构地区：[1]内蒙古农业大学乳品生物技术与工程教育部重点实验室,内蒙古呼和浩特010018

出　　处：《食品与生物技术学报》2009年第6期804-810,共7页Journal of Food Science and Biotechnology

基　　金：国家自然科学基金项目(30660135;30800861);国家863计划项目(2006AA10Z345;2007AA10Z353);教育部新世纪优秀人才支持计划项目(NCET-06-0269)

摘　　要：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。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 sequenees, 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.

关 键 词：乳杆菌 鉴定 限制性片段长度多态性(RFLP) 16S RDNA序列 

分 类 号：TS252.1[轻工技术与工程—农产品加工及贮藏工程]