新型克隆载体的构建与特性研究。III. 携带独特Eco RI位点的质粒pBR322衍生物，用于筛选由Eco RI产生的重组DNA分子。

Construction and characterization of new cloning vehicles. III. Derivatives of plasmid pBR322 carrying unique Eco RI sites for selection of Eco RI generated recombinant DNA molecules.

作者信息

Bolivar F

出版信息

Gene. 1978 Oct;4(2):121-36. doi: 10.1016/0378-1119(78)90025-2.

DOI:10.1016/0378-1119(78)90025-2

Abstract

In vitro recombinant DNA techniques were used to construct two new cloning vehicles, pBR324 and pBR235. 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 in order 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), Smal, (XmaI), BglII and DpnII restriction generated DNA molecules.

摘要

体外重组DNA技术被用于构建两种新的克隆载体，即pBR324和pBR235。这些载体源自质粒pBR322，是松弛型复制元件。质粒pBR324携带了pBR322中编码对氨苄青霉素（Apr）和四环素（Tcr）抗性的基因，以及源自质粒pMBI的大肠杆菌素E1结构基因和免疫基因。质粒pBR325携带了pBR322中的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分子中存在的唯一EcoRI限制性酶切位点位于大肠杆菌素E1结构基因（pBR324）或Cmr基因（pBR325）中。构建这些载体是为了使单个EcoRI位点位于一个结构基因的中间，当该结构基因失活时，便于选择质粒重组DNA分子。这些质粒允许对EcoRI、BamHI、HindIII、PstI、HincII、SalI、（XamI）、Smal、（XmaI）、BglII和DpnII限制性酶切产生的DNA分子进行分子克隆和轻松选择。