SHUTTLE VECTORS FOR THE ARCHAEBACTERIUM HALOBACTERIUM-VOLCANII
SHUTTLE VECTORS FOR THE ARCHAEBACTERIUM HALOBACTERIUM-VOLCANII
复制标题
DOI：
10.1073/pnas.86.14.5478
复制
发表时间：
1989-07-01
影响因子：
11.1
通讯作者：
DOOLITTLE, WF
中科院分区：
文献类型：
--
作者：
LAM, WL;DOOLITTLE, WF
考古细菌分子生物学的进步需要遗传分析工具。我们描述了可以在火山盐杆菌或大肠杆菌中选择和维护的载体。通过“鸟枪法”克隆内源火山嗜血杆菌内源质粒pHV2的衍生化产物，获得了抗性3-羟基-3-甲基戊二酰辅酶A还原酶抑制剂甲氧诺林的遗传决定基因pWL2，该基因可高频地将敏感的火山杆菌转化为甲氧草胺抗性。将抗性决定簇、部分PHV2和赋予氨苄西林和四环素抗性的pBR322衍生物pAT153连接在一起，形成穿梭载体pWL101和pWL102。我们描述了使用这些载体的条件，并初步定义了对耐药和对质粒复制和维护至关重要的区域。
Progress in archaebacterial molecular biology requires tools for genetic analysis. We describe vectors that can be selected and maintained in either Halobacterium volcanii or Escherichia coli. A genetic determinant for resistance to the 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor mevinolin was isolated by "shotgun cloning" into a derivative of the endogenous H. volcanii plasmid pHV2, to form pWL2, which transforms sensitive H. volcanii to mevinolin resistance at high frequency. The resistance determinant, portions of pHV2, and an ampicillin- and tetracycline-resistance-conferring pBR322 derivative, pAT153, were ligated together to form the shuttle vectors pWL101 and pWL102. We describe conditions for the use of these vectors and provide preliminary definition of regions essential for drug resistance and for plasmid replication and maintenance.