CONSTRUCTION AND CHARACTERIZATION OF NEW CLONING VEHICLES .3. DERIVATIVES OF PLASMID PBR322 CARRYING UNIQUE ECO RI SITES FOR SELECTION OF ECO RI GENERATED RECOMBINANT DNA-MOLECULES
CONSTRUCTION AND CHARACTERIZATION OF NEW CLONING VEHICLES .3. DERIVATIVES OF PLASMID PBR322 CARRYING UNIQUE ECO RI SITES FOR SELECTION OF ECO RI GENERATED RECOMBINANT DNA-MOLECULES
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DOI：
10.1016/0378-1119(78)90025-2
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发表时间：
1978-01-01
期刊：
影响因子：
3.5
通讯作者：
BOLIVAR, F
中科院分区：
文献类型：
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作者：
BOLIVAR, F
采用体外重组DNA技术构建了2种新型克隆载体pBR324和pBR325。这些源自质粒 pBR322 的载体是松弛复制元件。质粒pBR324携带来自pBR322的编码抗生素氨苄青霉素(Apr)和四环素(Tcr)抗性的基因以及源自质粒pMBI的大肠菌素E1结构和免疫基因。质粒pBR325携带来自pBR322的Apr和Tcr基因以及来自噬菌体P1Cm的氯霉素抗性基因(Cmr)。在这些质粒中，DNA 分子中存在的独特 EcoRI 限制位点位于大肠菌素 E1 结构基因 (pBR324) 或 Cmr 基因 (pBR325) 中。这些载体被构建为在结构基因的中间有一个单一的 EcoRI 位点，当该位点失活时，可以轻松选择质粒重组 DNA 分子。这些质粒允许分子克隆和轻松选择 EcoRI、BamHI、HindIII、PstI、HincII、SalI、(XamI)、SmaI、(XmaI)、BglII 和 DpnII 限制性生成的 DNA 分子。
In vitro recombinant DNA techniques were used to construct 2 new cloning vehicles, pBR324 and pBR325. These vectors, derived from plasmid pBR322, are relaxed replicating elements. Plasmid pBR324 carries the genes from pBR322 coding for resistance to the antibiotics ampicillin (Apr) and tetracycline (Tcr) and the colicin E1 structural and immunity genes derived from plasmid pMBI. Plasmid pBR325 carries the Apr and Tcr genes from pBR322 and the cloramphenicol resistance gene (Cmr) from phage P1Cm. In these plasmids the unique EcoRI restriction site present in the DNA molecule is located either in the colicin E1 structural gene (pBR324) or in the Cmr gene (pBR325). These vectors were constructed to have a single EcoRI site located in the middle of a structural gene which when inactivated would allow, for the easy selection of plasmid recombinant DNA molecules. These plasmids permit the molecular cloning and easy selection of EcoRI, BamHI, HindIII, PstI, HincII, SalI, (XamI), SmaI, (XmaI), BglII and DpnII restriction generated DNA molecules.