Barth C, Fraser D J, Fisher P R
School of Microbiology, La Trobe University, Bundoora 3083, Melbourne, Australia.
Nucleic Acids Res. 1998 Jul 1;26(13):3317-8. doi: 10.1093/nar/26.13.3317.
A rapid, simple method for characterization of plasmid insertions in the Dictyostelium discoideum genome was developed. It is based on the capability of linear plasmid multimers in the insertions to recircularize efficiently in Escherichia coli cells. This recombinational recircularization of plasmid multimers provides a highly sensitive and reliable tool for determining whether individual Dictyostelium transformants resulted from restriction enzyme-mediated integration (REMI) or from recombinational integration of plasmid (RIP). The method also reveals any rearrangements in RIP insertions and provides an estimate of the vector copy number in any particular transformant.
开发了一种快速、简单的方法来鉴定盘基网柄菌基因组中的质粒插入情况。该方法基于插入片段中的线性质粒多聚体在大肠杆菌细胞中高效环化的能力。质粒多聚体的这种重组环化提供了一种高度灵敏且可靠的工具，用于确定单个盘基网柄菌转化体是由限制酶介导的整合（REMI）产生的，还是由质粒的重组整合（RIP）产生的。该方法还能揭示RIP插入中的任何重排情况，并估算任何特定转化体中的载体拷贝数。
