Takeshita S, Sato M, Toba M, Masahashi W, Hashimoto-Gotoh T
Basic Research Department, Hoechst Japan Ltd, Kawagoe.
Gene. 1987;61(1):63-74. doi: 10.1016/0378-1119(87)90365-9.
Three types of alpha-complementation plasmid vectors were constructed which contain a chloramphenicol- or kanamycin-resistance (CmR or KmR) gene and polylinker cloning sites within the coding region of lacZ'. These vectors are essentially based on high- or low-copy-number replicons. The low-copy-number vectors, 3.61 kb in size, confer CmR and contain the pSC101 replicon and pUC8-/pUC9-type polylinker. On the other hand, the high-copy-number vectors, 2.21 to 2.68 kb in size, confer either CmR or KmR, and contain the pBR322 replicon and pUC18-/pUC19-type or other modified polylinkers. All cloning sites except HindIII and SmaI sites in the KmR vectors are unique in each plasmid. Since almost all frequently used plasmid vectors confer ampicillin resistance, these vectors may be useful to simplify the subcloning/DNA joining experiments due to unnecessity of radioisotope labelling, size fractionation and purification of foreign DNA segments.
构建了三种α-互补质粒载体，它们在lacZ'编码区内含有氯霉素或卡那霉素抗性（CmR或KmR）基因以及多克隆位点克隆位点。这些载体基本上基于高拷贝数或低拷贝数复制子。低拷贝数载体大小为3.61 kb，赋予CmR抗性，含有pSC101复制子和pUC8-/pUC9型多克隆位点。另一方面，高拷贝数载体大小为2.21至2.68 kb，赋予CmR或KmR抗性，含有pBR322复制子和pUC18-/pUC19型或其他修饰的多克隆位点。除KmR载体中的HindIII和SmaI位点外，每个质粒中的所有克隆位点都是唯一的。由于几乎所有常用的质粒载体都赋予氨苄青霉素抗性，这些载体可能有助于简化亚克隆/DNA连接实验，因为无需进行放射性同位素标记、大小分级分离和外源DNA片段的纯化。
