两株假单胞菌菌株的RAPD分析及分子生物学鉴定  

RAPD Analysis and Molecule Biological Identification of two Strains of Acinetobacter calcoaceticus

作　　者：黄玲[1] 夏立秋[1] 刘萍[1] 周卫东 

机构地区：[1]湖南师范大学生命科学学院,中国湖南长沙410081 [2]北京鼎国生物技术发展中心,中国北京100088

出　　处：《生命科学研究》2006年第S2期78-84,共7页Life Science Research

基　　金：国家自然科学基金项目资助(30570050)

摘　　要：根据DNA随机扩增多态性(RandomAmplifiedPolymorphicDNA,简RAPD)分子标记技术设计鉴别引物,建立一种快速、准确检测病人体内新发现的假单胞菌菌株的分子生物学方法.采用RAPD分析方法对该菌种的对照菌株AcinetobactercalcoaceticusKHW14(简称A.calcoaceticusKHW14)和新分离的菌株Acinetobactercalcoaceticus(简称A.calcoaceticus)进行指纹分析,依据两菌株的差异序列设计两对引物,并建立最佳的PCR扩增体系,产物经1.2%琼脂糖凝胶电泳得菌株特异性电泳图谱.此图谱可作为鉴定两菌株的标准图谱,RAPD分析方法具有良好的重复性,同时也进一步验证了两菌株的同源性.A rapid and sensitive molecular method which could detect a certain bacterium isolated in the body of patient is established according to the random amplified polymorphic DNA(RAPD) analysis.In order to distinguish the two different bacterium isolates,the genomic DNA of the comparison strain(Acinetobacter calcoaceticus KHW14) and newly isolated strain(Acinetobacter calcoaceticus) were extracted at first and then analyzed through PCR-based RAPD fingerprinting.According to the different sequence gotten from RAPD method two couples of primers were designed.The PCR system was established with genomic DNA as templates.The strain-characteristic gel electrophoreses profiles could be used as the marker for rapidly and exactly distinguishing,detecting the two bacteria strains.

关 键 词：细菌 RAPD分析 分子生物学鉴定 菌株特异性图谱 

分 类 号：Q93-331[生物学—微生物学]