庆大霉素耐药基因的分子克隆  

MOLECULAR CLONING OF GENTAMICIN RESISTANCE GENE

作　　者：严世方[1] 包幼迪[1] 

机构地区：[1]福建医学院基因工程研究室,福州350004

出　　处：《中国抗生素杂志》1990年第5期333-338,共6页Chinese Journal of Antibiotics

摘　　要：利用基因工程技术,从一株流行的致病性大肠杆菌的65Md的可传递的多重耐药质粒pEFM2出发,克隆获得一重组质粒pBY101,它仅表达庆大霉素和四环素抗性,其庆大霉素耐药基因在8.9kb的Pst Ⅰ片段上。再由EcoRⅠ酶解pBY101,亚克隆获得一重组质粒pBY 102,其庆大霉素耐药基因在4.9kb的Pst Ⅰ-EcoRⅠ片段上。经氨基糖苷类抗生素底物谱分析和用生物素标记的2″-O-腺苷转移酶[ANT(2″)]基因探针,以Southern印迹杂交法检测,证明庆大霉素耐药基因不是编码ANT(2″)的。A 8.9-kb Pst I fragment was cloned from the transconjugant of a clinical isolat of epidemic enteropathogenic Escherichia coli (EPEC) containing a 65 Md multiresistance Plasmid pEFM2. The recombinant plasmid pBY101 conferred resistance to gentamicin and tetracycline. This clone was used to generate a series of subclones from which a 4.9-kb PstI-EcoRI fragment containing a gentamicin resistance gene was obtained. The recombinant plasmid was named pBYl02. On the basis of aminoglycoside resistance patterns it appeared that the amiaoglycoside-modifying enzyme was not a 2'-O-adenyltransferaseCANT(2')]. This was tested and confirmed by Southern hybridization with a biotia-labeled 2.0-kb BamH Ⅰ-Hind Ⅲ probe of pDG0103 containing a gentamicin ANT(2')geng.

关 键 词：庆大霉素 耐药基因 分子克隆 

分 类 号：TQ465.2[化学工程—制药化工]