Meijer W J, Lewis P J, Errington J, Salas M
Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Universidad Autónoma, Canto Blanco, 28049 Madrid, Spain.
EMBO J. 2000 Aug 1;19(15):4182-90. doi: 10.1093/emboj/19.15.4182.
We have examined the localization of DNA replication of the Bacillus subtilis phage phi 29 by immunofluorescence. To determine where phage replication was localized within infected cells, we examined the distribution of phage replication proteins and the sites of incorporation of nucleotide analogues into phage DNA. On initiation of replication, the phage DNA localized to a single focus within the cell, nearly always towards one end of the host cell nucleoid. At later stages of the infection cycle, phage replication was found to have redistributed to multiple sites around the periphery of the nucleoid, just under the cell membrane. Towards the end of the cycle, phage DNA was once again redistributed to become located within the bulk of the nucleoid. Efficient redistribution of replicating phage DNA from the initial replication site to various sites surrounding the nucleoid was found to be dependent on the phage protein p16.7.
我们通过免疫荧光法检测了枯草芽孢杆菌噬菌体phi 29的DNA复制定位。为了确定噬菌体复制在受感染细胞内的定位位置,我们检测了噬菌体复制蛋白的分布以及核苷酸类似物掺入噬菌体DNA的位点。在复制起始时,噬菌体DNA定位于细胞内的一个单一焦点,几乎总是朝向宿主细胞核的一端。在感染周期的后期,发现噬菌体复制重新分布到核周周围的多个位点,就在细胞膜下方。在周期结束时,噬菌体DNA再次重新分布,位于核的大部分区域内。发现复制中的噬菌体DNA从初始复制位点有效地重新分布到核周周围的各个位点取决于噬菌体蛋白p16.7。