Seed B
Nucleic Acids Res. 1983 Apr 25;11(8):2427-45. doi: 10.1093/nar/11.8.2427.
Cloned genes have been purified from recombinant DNA bacteriophage libraries by a method exploiting homologous reciprocal recombination in vivo. In this method 'probe' sequences are inserted in a very small plasmid vector and introduced into recombination-proficient bacterial cells. Genomic bacteriophage libraries are propagated on the cells, and phage bearing sequences homologous to the probe acquire an integrated copy of the plasmid by reciprocal recombination. Phage bearing integrated plasmids can be purified from the larger pool of phage lacking plasmid integrates by growth under the appropriate selective conditions.
已通过一种利用体内同源性相互重组的方法从重组DNA噬菌体文库中纯化出克隆基因。在这种方法中,“探针”序列被插入一个非常小的质粒载体中,并导入具有重组能力的细菌细胞。基因组噬菌体文库在这些细胞中增殖,携带与探针同源序列的噬菌体通过相互重组获得质粒的整合拷贝。通过在适当的选择条件下培养,可以从缺乏质粒整合体的较大噬菌体群体中纯化出携带整合质粒的噬菌体。