Barreiro V, Haggård-Ljungquist E
Department of Microbial Genetics, Karolinska Institutet, Stockholm, Sweden.
J Bacteriol. 1992 Jun;174(12):4086-93. doi: 10.1128/jb.174.12.4086-4093.1992.
Integration of bacteriophage P2 into the Escherichia coli genome involves recombination between two attachment sites, attP and attB, one on the phage and one on the host genome, respectively. At least 10 different attB sites have been identified over the years. In E. coli C, one site, called locI, is preferred, being occupied before any of the others. In E. coli K-12, no such preference is seen (reviewed in L. E. Bertani and E. W. Six, p. 73-143, in R. Calendar, ed., The Bacteriophages, vol. 2, 1988). The DNA sequence of locI has been determined, and it shows a core sequence of 27 nucleotides identical to attP (A. Yu, L. E. Bertani, and E. Haggård-Ljungquist, Gene 80:1-12, 1989). By inverse polymerase chain reactions, the prophage-host junctions of DNA extracted from P2 lysogenic strains have been amplified, cloned, and sequenced. By combining the attL and attR sequences, the attB sequences of locations II, III, and H have been deduced. The core sequence of location II had 20 matches to the 27-nucleotide core sequence of attP; the sequences of locations III and H had 17 matches. Thus, the P2 integrase accepts at least up to 37% mismatches within the core sequence. The E. coli K-12 strains examined all contain a 639-nucleotide-long cryptic remnant of P2 at a site with a sequence similar to that of locI but that may have a different map position. The P2 remnant consists of the C-terminal part of gene D, all of gene ogr, and attR. Locations II, III, and H have been located on Kohara's physical map to positions 3670, 1570 to 1575, and 2085, respectively.
噬菌体P2整合到大肠杆菌基因组中涉及两个附着位点attP和attB之间的重组,这两个位点分别位于噬菌体和宿主基因组上。多年来已鉴定出至少10个不同的attB位点。在大肠杆菌C中,一个名为locI的位点是首选,它会在其他位点之前被占据。在大肠杆菌K-12中,则没有这种偏好(见L. E. 贝尔塔尼和E. W. 西克斯所著,第73 - 143页,载于R. 卡尔日历编著的《噬菌体》第2卷,1988年)。已确定locI的DNA序列,其显示出与attP相同的27个核苷酸的核心序列(A. 于、L. E. 贝尔塔尼和E. 哈加德 - 伦奎斯特,《基因》80:1 - 12,1989年)。通过反向聚合酶链反应,从P2溶原性菌株中提取的DNA的原噬菌体 - 宿主连接点已被扩增、克隆和测序。通过合并attL和attR序列,推导出了位置II、III和H的attB序列。位置II的核心序列与attP的27个核苷酸核心序列有20个匹配;位置III和H的序列有17个匹配。因此,P2整合酶在核心序列内可接受至少高达37%的错配。所检测的大肠杆菌K-12菌株在一个位点都含有一段639个核苷酸长的P2隐匿残余片段,该位点的序列与locI相似,但可能具有不同的图谱位置。P2残余片段由基因D的C末端部分、整个基因ogr和attR组成。位置II、III和H已在小原的物理图谱上分别定位到3670、1570至1575以及2085的位置。