Summers W C, Glazer P M, Malkevich D
Radiobiology Laboratories, Yale University School of Medicine, New Haven, CT 06510.
Mutat Res. 1989 Mar-May;220(2-3):263-8. doi: 10.1016/0165-1110(89)90030-4.
Foreign DNA sequences contained in lambda bacteriophage genomes integrated in mammalian DNA can be efficiently rescued into infectious phage particles by treatment of the mammalian DNA with lambda-packaging extracts prepared in E. coli. This system provides for rapid, non-selective recovery of stably integrated, chromosomal sequences into lambda phage for subsequent analysis in bacterial systems. Since rescue is prior to selection, mutations can be recovered from intact animals made transgenic for the phage-target gene sequences. Such approaches allow study of physiologically relevant aspects of mammalian mutagenesis at the molecular level.
整合于哺乳动物DNA中的λ噬菌体基因组所含的外源DNA序列,通过用在大肠杆菌中制备的λ包装提取物处理哺乳动物DNA,能够高效地拯救到感染性噬菌体颗粒中。该系统可将稳定整合的染色体序列快速、非选择性地拯救到λ噬菌体中,以便在细菌系统中进行后续分析。由于拯救先于筛选,因此可以从针对噬菌体靶基因序列制作的完整转基因动物中回收突变。这些方法能够在分子水平上研究哺乳动物诱变的生理相关方面。