Windle B E
Gene. 1986;45(1):95-9. doi: 10.1016/0378-1119(86)90136-8.
Two new lambda vectors were constructed which permit cloning of genes that are potentially lethal if cloned in analogous plasmid vectors. lambda DL10 and lambda DL11 contain the alpha-complementing fragment of lacZ and multiple cloning sites found in the polylinker region of M13mp10 and M13mp11, respectively. DNA cloned into the unique cloning sites of these vectors can be detected by inactivation of alpha-complementation. These lambda vectors provide a lac promoter for expression of foreign genes as well as the ability to make fusion proteins. Two plasmid expression vectors, pPR110 and pPR111, were constructed from lambda DL10 and lambda DL11 respectively, and pCQV2. These plasmids, which express lacZ alpha-complementing activity from the lambda PR promoter, contain multiple cloning sites immediately downstream of the PR promoter. They allow cloning of genes under the control of the PR promoter and the plasmid-encoded thermosensitive (cI857) repressor, and allow easy detection of inserted fragments by inactivation of alpha-complementation.
构建了两种新的λ噬菌体载体,它们能够克隆那些若克隆到类似质粒载体中可能具有致死性的基因。λ DL10和λ DL11分别含有lacZ的α-互补片段以及在M13mp10和M13mp11的多克隆位点区域中发现的多克隆位点。克隆到这些载体独特克隆位点的DNA可通过α-互补的失活来检测。这些λ噬菌体载体为外源基因的表达提供了一个lac启动子,同时具备产生融合蛋白的能力。分别从λ DL10和λ DL11以及pCQV2构建了两种质粒表达载体pPR110和pPR111。这些质粒从λ PR启动子表达lacZα-互补活性,在PR启动子下游紧邻处含有多克隆位点。它们允许在PR启动子和质粒编码的温度敏感型(cI857)阻遏物的控制下克隆基因,并通过α-互补的失活轻松检测插入片段。