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一个含有乳链菌肽抗性基因的乳酸乳球菌质粒pTS50的鉴定
引用本文:汤莎,陈秀珠,杨巍,陈美玲,还连栋.一个含有乳链菌肽抗性基因的乳酸乳球菌质粒pTS50的鉴定[J].微生物学报,2001,41(5):536-541.
作者姓名:汤莎  陈秀珠  杨巍  陈美玲  还连栋
作者单位:中国科学院微生物研究所,
基金项目:中国科学院重点研究项目(KY95-J1-331)
摘    要:在添加乳链菌肽、乳糖及溴甲酚紫的M1 7选择培养基上 ,从 1 97个新鲜牛奶样品中筛选到 3株乳链菌肽抗性菌株 ,PCR扩增证实它们都含有乳链菌肽抗性基因。菌种生理生化特性鉴定及特异性 1 6SrDNAPCR扩增产物的序列测定结果表明这 3株菌都属于乳酸乳球菌乳酸亚种。质粒转化实验发现乳酸乳球菌乳酸亚种TS 1 640中的乳链菌肽抗性基因位于一个约47kb的大质粒pTS50上。BamHI、EcoRI、HindⅢ、NcoI、PstⅠ酶切分析和Southern杂交 ,进一步将乳链菌肽抗性基因定位于pTS50的一个约 1 9kbEcoRI酶切片段中

关 键 词:乳酸乳球菌    乳链菌肽抗性    质粒    酶切分析    Southern杂交
文章编号:0001-6209(2001)05-0536-06
修稿时间:2000年11月15

ISOLATION AND CHARACTERIZATION OF A PLASMID pTS50, WHICH ENCODES NISIN RESISTANCE DETERMINANT IN LACTOCOCCUS LACTIS TS1640
Tang Sha\ Chen Xiuzhu\ Yang Wei\ Chen Meiling\ Huan Liandong.ISOLATION AND CHARACTERIZATION OF A PLASMID pTS50, WHICH ENCODES NISIN RESISTANCE DETERMINANT IN LACTOCOCCUS LACTIS TS1640[J].Acta Microbiologica Sinica,2001,41(5):536-541.
Authors:Tang Sha\ Chen Xiuzhu\ Yang Wei\ Chen Meiling\ Huan Liandong
Institution:Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China.
Abstract:Three nisin-resistant Lactococcus lactis strains were screened from 197 samples of fresh milk on a selective medium (M17) supplemented with nisin, lactose and bromocresol purple, and were confirmed to have the nisin resistance determinant (nsr) by PCR amplification. Physiological and biochemical tests as well as Lactococcus lactis specific 16S rDNA sequence analysis revealed that the three strains all belong to Lactococcus lactis subsp. lactis. A large plasmid, pTS50, was identified in L. lactis subsp. lactis TS1640 which encodes resistance to nisin following electroporation of the total plasmids DNA into L. lactis MG1363. The molecular weight of plasmid pTS50 was estimated to be 47 kb by restriction analysis of BamHI, EcoRI, HindIII, NcoI, PstI and nsr was localized on a 1.9 kb EcoRI fragment by Southern hybridization.
Keywords:Lactococcus lactis    Nisin resistance  Plasmid  Restriction enzyme digest  Southern blot
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