Gambill B D, Summers A O
Gene. 1985;39(2-3):293-7. doi: 10.1016/0378-1119(85)90326-9.
Cloning vectors have been constructed employing two diverse replicons, IncQ and P15A. Both vectors confer resistance to kanamycin (Km) and mercuric ions (Hg2+). One of these vectors, pDG105, is a broad-host-range, nonconjugative, oligocopy IncQ plasmid, which is capable of transforming Escherichia coli, Acinetobacter calcoaceticus, and Pseudomonas putida. The second vector, pDG106, is a narrow-host-range, multicopy cloning vector compatible with pBR322. Both vectors contain unique cloning sites in the Km-resistance gene for HindIII, SmaI, and XhoI, as well as unique EcoRI and ScaI sites in the mer operon. Cloning into the EcoRI site in the mer operon results in the mercury "supersensitive" phenotype, easily detectable by replica plating. Insertion of the galK gene into the EcoRI site in the mer operon results in Hg2+-inducible galactokinase activity, demonstrating the application of these plasmids as regulated expression vectors.
已构建了采用两种不同复制子（IncQ和P15A）的克隆载体。这两种载体都赋予对卡那霉素（Km）和汞离子（Hg2+）的抗性。其中一种载体pDG105是一种广宿主范围、非接合型、低拷贝的IncQ质粒，能够转化大肠杆菌、乙酸钙不动杆菌和恶臭假单胞菌。第二种载体pDG106是一种窄宿主范围、多拷贝的克隆载体，与pBR322兼容。两种载体在Km抗性基因中都含有用于HindIII、SmaI和XhoI的独特克隆位点，以及在汞操纵子中含有独特的EcoRI和ScaI位点。克隆到汞操纵子中的EcoRI位点会导致汞“超敏感”表型，通过影印平板法很容易检测到。将galK基因插入汞操纵子中的EcoRI位点会导致Hg2+诱导的半乳糖激酶活性，证明了这些质粒作为调控表达载体的应用。
