Boyd A C, Sherratt D J
MRC Human Genetics Unit, Western General Hospital, Edinburgh, UK.
Gene. 1995 Feb 3;153(1):57-62. doi: 10.1016/0378-1119(94)00788-t.
A series of general-purpose plasmid vectors based on the phage lambda origin of replication (ori) has been constructed. Each vector consists of a backbone plasmid encoding chloramphenicol resistance (CmR) and containing a unique HaeII site into which the lacZ alpha-complementing multiple cloning site (MCS) region of an established vector was inserted. To increase the cloning potential of the inserted MCS, superfluous restriction sites in the backbone were removed by a variety of techniques. The vectors, designated pCLIP (for CmR lambda ori integration proficient) plasmids, are of medium copy number and are compatible with most other vectors in common use. A total of 17 unique restriction sites in pCLIP8, pCLIP9, pCLIP18, pCLIP19 and pCLIP23 are available for cloning. As well as possessing the usual properties of vectors, the pCLIP plasmids are able to integrate reversibly into lambda prophage by homologous recombination. Thus, cloned DNA can be maintained in single or multiple copy at will. By integrating recombinant plasmids into appropriate deletion prophages followed by induction, phage::plasmid hybrids are produced which can be manipulated as phage. These properties are demonstrated using a test recombinant plasmid, pCLIPLEU2. The pCLIP vectors are therefore useful for routine plasmid cloning, complementation analysis and applications where the ability to manipulate recombinants in plasmid, phage or prophage forms is advantageous.
已构建了一系列基于噬菌体λ复制起点（ori）的通用质粒载体。每个载体都包含一个编码氯霉素抗性（CmR）的主体质粒，并含有一个独特的HaeII位点，已将一个成熟载体的lacZα互补多克隆位点（MCS）区域插入该位点。为了提高插入的MCS的克隆潜力，通过多种技术去除了主体质粒中的多余限制酶切位点。这些载体被命名为pCLIP（意为氯霉素抗性λ ori整合 proficient）质粒，属于中等拷贝数，并且与大多数其他常用载体兼容。pCLIP8、pCLIP9、pCLIP18、pCLIP19和pCLIP23共有17个独特的限制酶切位点可用于克隆。除了具备载体的常规特性外，pCLIP质粒还能够通过同源重组可逆地整合到λ原噬菌体中。因此，克隆的DNA可以随意以单拷贝或多拷贝形式维持。通过将重组质粒整合到适当的缺失原噬菌体中，然后进行诱导，可产生噬菌体::质粒杂种，其可作为噬菌体进行操作。使用测试重组质粒pCLIPLEU2证明了这些特性。因此，pCLIP载体可用于常规质粒克隆、互补分析以及在质粒、噬菌体或原噬菌体形式中操作重组体具有优势的应用。
