Chaconas G, de Bruijn F J, Casadaban M J, Lupski J R, Kwoh T J, Harshey R M, DuBow M S, Bukhari A I
Gene. 1981 Jan-Feb;13(1):37-46. doi: 10.1016/0378-1119(81)90041-x.
Recombinant plasmids carrying one or both ends of the bacteriophage Mu genome were constructed by molecular cloning. Transposable mini-Mu's with selectable markers (ampicillin resistance, kanamycin resistance or the entire lac operon of Escherichia coli) inserted between the Mu ends were also constructed. As a source of lac operon DNA, a pBR322 derivative with a 27 kb insert containing the lac operon was constructed. The plasmids with both ends of Mu (mini-Mu's) conferred full Mu immunity upon the host cells. However, the same mini-Mu's containing kan or lac inserts were defective in immunity. A summary of the construction and physical characterization, including restriction endonuclease cleavage maps and some of the biological properties of the plasmids, is presented.
通过分子克隆构建了携带噬菌体Mu基因组一端或两端的重组质粒。还构建了在Mu两端之间插入了选择标记(氨苄青霉素抗性、卡那霉素抗性或大肠杆菌完整的乳糖操纵子)的转座型小Mu。作为乳糖操纵子DNA的来源,构建了一种带有包含乳糖操纵子的27 kb插入片段的pBR322衍生物。带有Mu两端的质粒(小Mu)赋予宿主细胞完全的Mu免疫性。然而,含有卡那或乳糖插入片段的相同小Mu在免疫性方面存在缺陷。本文介绍了这些质粒的构建和物理特性总结,包括限制性内切酶切割图谱以及一些生物学特性。