van den Berg E A, Geerse R H, Memelink J, Bovenberg R A, Magnée F A, van de Putte P
Nucleic Acids Res. 1985 Mar 25;13(6):1829-40. doi: 10.1093/nar/13.6.1829.
A region located upstream of the uvrB promoters P1 and P2 was found to cause high plasmid loss when cloned in multicopy vectors. Two sequence elements responsible for this phenomenon were identified by mapping of spontaneous mutations that restore plasmid maintenance: a sequence known to have in vitro promoter activity and a partially overlapping sequence that shows extensive homology to recognition sites for the DnaA protein. Accordingly alterations in the level of DnaA protein in vivo were found to affect the extent of plasmid loss. A possible role for interaction of the DnaA protein with the region of interest is discussed in relation to regulation of uvrB expression.
当克隆到多拷贝载体中时,uvrB启动子P1和P2上游的一个区域会导致高频率的质粒丢失。通过对恢复质粒维持的自发突变进行定位,确定了导致这种现象的两个序列元件:一个已知具有体外启动子活性的序列,以及一个与DnaA蛋白识别位点具有广泛同源性的部分重叠序列。因此,发现体内DnaA蛋白水平的改变会影响质粒丢失的程度。结合uvrB表达的调控,讨论了DnaA蛋白与相关区域相互作用的可能作用。