Brandis Gerrit, Cao Sha, Hughes Diarmaid
Department of Medical Biochemistry and Microbiology, Box 582 Biomedical Center, Uppsala University, Uppsala, Sweden.
Bio Protoc. 2019 Feb 5;9(3):e3159. doi: 10.21769/BioProtoc.3159.
Homologous recombination between two similar DNA molecules, plays an important role in the repair of double-stranded DNA breaks. Recombination can occur between two sister chromosomes, or between two locations of similar sequence identity within the same chromosome. The assay described here is designed to measure the rate of homologous recombination between two locations with sequence similarity within the same bacterial chromosome. For this purpose, a selectable/counter-selectable genetic cassette is inserted into one of the locations and homologous recombination repair rates are measured as a function of recombinational removal of the inserted cassette. This recombinational repair process is called gene conversion, non-reciprocal recombination. We used this method to measure the recombination rates between genes within gene families and to study the stability of mobile genetic elements inserted into members of gene families.
两个相似DNA分子之间的同源重组在双链DNA断裂修复中起重要作用。重组可发生在两条姐妹染色体之间，或同一染色体上两个具有相似序列同一性的位置之间。此处描述的测定方法旨在测量同一细菌染色体内两个具有序列相似性的位置之间的同源重组率。为此，将一个可选择/反选择遗传盒插入其中一个位置，并根据插入盒的重组去除情况来测量同源重组修复率。这种重组修复过程称为基因转换，即非相互重组。我们使用这种方法来测量基因家族内基因之间的重组率，并研究插入基因家族成员中的移动遗传元件的稳定性。
