Sandmeier H, Iida S, Hübner P, Hiestand-Nauer R, Arber W
Abteilung Mikrobiologie, Universität Basel, Switzerland.
Nucleic Acids Res. 1991 Nov 11;19(21):5831-8. doi: 10.1093/nar/19.21.5831.
The bacteriophage P1-related plasmid p15B of E. coli 15T- contains a 3.5 kb long region which frequently undergoes complex rearrangements by DNA inversion. Site-specific recombination mediated by the Min DNA invertase occurs at six crossover sites and it eventually results in a population of 240 isomeric configurations of this region. We have determined 8.3-kb sequences of the invertible DNA and its flanking regions. The result explains how DNA inversion fuses variable 3' parts to a constant 5' part, thereby alternatively assembling one out of six different open reading frames (ORF). The resulting variable gene has a coding capacity of between 739 and 762 amino acids. A large portion of its constant part is composed of repeated sequences. The p15B sequences in front of the variable fusion gene encode a small ORF and a phage-specific late promoter and are highly homologous to P1 DNA. Adjacent to the DNA invertase gene min, we have found a truncated 5' region of a DNA invertase gene termed psi cin which is highly homologous to the phage P1 cin gene. Its recombinational enhancer segment is inactive, but it can be activated by the substitution of two nucleotides.
大肠杆菌15T - 的噬菌体P1相关质粒p15B含有一个3.5 kb长的区域,该区域经常通过DNA倒位发生复杂的重排。由Min DNA转化酶介导的位点特异性重组发生在六个交叉位点,最终导致该区域出现240种异构构型。我们已经确定了可倒位DNA及其侧翼区域的8.3 kb序列。结果解释了DNA倒位如何将可变的3'部分与恒定的5'部分融合,从而从六个不同的开放阅读框(ORF)中交替组装出一个。产生的可变基因编码能力在739至762个氨基酸之间。其恒定部分的很大一部分由重复序列组成。可变融合基因前面的p15B序列编码一个小ORF和一个噬菌体特异性晚期启动子,并且与P1 DNA高度同源。在DNA转化酶基因min旁边,我们发现了一个称为psi cin的DNA转化酶基因的截短5'区域,它与噬菌体P1 cin基因高度同源。其重组增强子片段是无活性的,但可以通过两个核苷酸的取代而被激活。