耐氟菌株及其亲代菌株ATP酶基因的DNA测序  

ATPase Gene DNA Sequence Analysis of Fluoride-Resistant Strains and Their Parent Strains

作　　者：盛江筠[1] 朱敏[1] 黄正蔚[1] 刘正[1] 

机构地区：[1]上海第二医科大学口腔医学院第九人民医院口腔内科,上海200011

出　　处：《上海第二医科大学学报》2005年第4期340-341,共2页Acta Universitatis Medicinalis Secondae Shanghai

基　　金：国家自然科学基金(39870780)资助项目.

摘　　要：目的对变型链球菌耐氟菌株及其亲代菌株的胞膜ATP酶基因进行DNA序列分析。方法分别制备变链菌株Ingbritt及其耐氟菌株IngbrittFR的菌悬液(OD600nm=1.0),经溶菌酶、Lysis消化。按已知ATP酶基因引物设计、扩增ATP酶各片段。将ATP酶基因连接至PUCmT克隆载体,转化后经限制性内切酶酶切鉴定为阳性克隆。采用DG1G1E检测ATP酶基因,DNA测序试剂盒分析DNA序列。结果耐氟菌株与其亲代菌株的核苷酸序列无显著差别。结论ATP酶基因改变不是变链菌株耐氟突变的机制。Objective In order to demonstrate the idiotypic variation between fluoride-resistant strains and their pa- rent strains, ATPase gene sequence analysis was carried out. Methods The suspensions of S.mutans Ingbritt and their fluoride-resistant strains, Ingbritt-FR, were prepared, respectively, and the bacteria were digested with lysozyme and lysis. The fragments of ATPase gene were amplified according to the primer design of ATPase before transformation,the ATPase gene was connected with PUCm-T clone carrier, the plasmid DNA transformation were identified with restriction endoneuclease and confirmed by DG 1G 1E assay. The DNA sequence was analyzed. Results There was no significant difference in necleotide sequence between fluoride-resistant strains and their parent strains. Conclusion The mechanism of fluoride-resistant strain may not be the result of the variation of ATPase gene.

关 键 词：变链菌 耐氟菌株 ATP酶基因 DNA序列 

分 类 号：R446.5[医药卫生—诊断学]