Pato M L, Howe M M, Higgins N P
Department of Microbiology and Immunology, University of Colorado Health Sciences Center, Denver 80262.
Proc Natl Acad Sci U S A. 1990 Nov;87(22):8716-20. doi: 10.1073/pnas.87.22.8716.
We have discovered a centrally located site that is required for efficient replication of bacteriophage Mu DNA and identified it as a strong DNA gyrase-binding site. Incubation of Mu DNA with gyrase and enoxacin revealed a cleavage site 18.1 kilobases from the left end of the 37.2-kilobase genome. Two observations indicate a role for the site in Mu replication: mutants of Mu, able to grow on an Escherichia coli gyrB host that does not allow growth of wild-type Mu, were found to possess single-base changes resulting in more efficient gyrase binding and cleavage at the site. Introduction of a 147-base-pair deletion that eliminated the site from a prophage inhibited the onset of Mu replication for greater than 1 hr after induction.
我们发现了一个噬菌体Mu DNA高效复制所需的位于中心位置的位点,并将其鉴定为一个强DNA促旋酶结合位点。将Mu DNA与促旋酶和依诺沙星一起温育,揭示了一个位于37.2千碱基基因组左端18.1千碱基处的切割位点。两项观察结果表明该位点在Mu复制中起作用:发现能够在不允许野生型Mu生长的大肠杆菌gyrB宿主上生长的Mu突变体,具有单碱基变化,导致在该位点更有效地结合和切割促旋酶。引入一个147碱基对的缺失,该缺失从原噬菌体中消除了该位点,抑制了诱导后超过1小时的Mu复制起始。