Liljeström P, Pirhonen M, Palva E T
Gene. 1985;40(2-3):241-6. doi: 10.1016/0378-1119(85)90046-0.
A technique is presented by which chromosomal mutations may be efficiently transferred onto chimeric multicopy plasmids in vivo. The technique employs the transduction of plasmids using bacteriophage P1 as vector. The utility of this method was demonstrated by cloning a chromosomal ompR mutation of Escherichia coli K-12. The high-frequency transduction of the chimeric plasmid appeared to be dependent on its integration into the chromosome by homologous recombination. The results also suggest that the plasmid was transduced as part of the chromosome and resolved from its integrated state in the recipient cell, resulting in a high yield of mutant plasmid segregants.
本文介绍了一种可在体内将染色体突变高效转移至嵌合多拷贝质粒的技术。该技术利用噬菌体P1作为载体转导质粒。通过克隆大肠杆菌K-12的染色体ompR突变证明了此方法的实用性。嵌合质粒的高频转导似乎依赖于其通过同源重组整合到染色体中。结果还表明，质粒作为染色体的一部分被转导，并在受体细胞中从其整合状态解离，从而产生高产率的突变体质粒分离物。
