Cloning, isolation, and characterization of replication regions of complex plasmid genomes.
Cloning, isolation, and characterization of replication regions of complex plasmid genomes.
复制标题
复杂质粒基因组复制区域的克隆、分离和表征。
DOI：
10.1073/pnas.72.6.2242
复制
发表时间：
1975
影响因子：
11.1
通讯作者：
S. Cohen
中科院分区：
文献类型：
--
作者：
K. Timmis;F. Cabello;S. Cohen
EcoRI 核酸内切酶生成的携带 F'lac 和 R6-5 质粒复制区的 DNA 片段已被克隆和分离，使用源自金黄色葡萄球菌质粒的非复制氨苄青霉素抗性 DNA 片段作为选择载体。对构建的质粒嵌合体和亲本复制子进行异源双链分析，将克隆的 R6-5 复制区定位到 R6-5 图谱上千碱基对坐标 1.0 和 88.0 之间的 DNA 片段。两个亲本复制子的质粒复制功能和控制质粒不相容性的基因座之间的物理接近性已被证明。报道的克隆方法似乎普遍适用于识别和分离各种复杂基因组的复制区域。
EcoRI endonuclease-generated DNA fragments carrying replication regions of the F'lac and R6-5 plasmids have been cloned and isolated, using as a selection vehicle a nonreplicating ampicillin-resistance DNA fragment derived from a Staphylococcus aureus plasmid. Heteroduplex analysis of the constructed plasmid chimeras and the parent replicons has localized the cloned R6-5 replication region to a DNA segment between kilobase pair coordinates 1.0 and 88.0 on the R6-5 map. Physical proximity between the plasmid replication functions and the locus governing plasmid incompatibility has been shown for both parent replicons. The cloning method reported appears to be generally applicable for the identification and isolation of replication regions of a variety of complex genomes.