Dept. of Genetics, Microbiology and Toxicology, Stockholm University, Stockholm, Sweden.
Virology. 2010 Dec 5;408(1):64-70. doi: 10.1016/j.virol.2010.08.035. Epub 2010 Sep 28.
Phage integrases have the potential of becoming tools for safe site-specific integration of genes into unmodified human genomes. The P2-like phages have been found to have different bacterial host integration sites and consequently they have related integrases with different sequence specificities. In this work the site-specific recombination system of the P2-like phage ΦD145 is characterized. The minimal attB site is determined to 22 nt with 18 nt identity to the core region of attP. A non-coding sequence on the human chromosome 13 is shown to be a rather good substrate for recombination in vivo in bacteria as well as in a plasmid system in HeLa cells when HMG protein recognition sequences are inserted between the left arm-binding site and the core in the complex phage attachment site attP. Thus ΦD145 integrase that belongs to the tyrosine family shows potential as a tool for site-specific integration into the human genome.
噬菌体整合酶有可能成为将基因安全且特异性地整合到未经修饰的人类基因组中的工具。已发现 P2 样噬菌体具有不同的细菌宿主整合位点，因此它们具有相关的整合酶，具有不同的序列特异性。在这项工作中，对 P2 样噬菌体 ΦD145 的特异性重组系统进行了表征。最小的 attB 位点被确定为 22 个核苷酸，与 attP 的核心区域有 18 个核苷酸的同一性。在细菌以及在含有 HMG 蛋白识别序列的 HeLa 细胞质粒系统中，当在复杂的噬菌体附着位点 attP 中，在左臂结合位点和核心之间插入非编码序列时，人类染色体 13 上的一个非编码序列显示出在体内进行重组的良好底物。因此，属于酪氨酸家族的 ΦD145 整合酶有可能成为人类基因组特异性整合的工具。
