Medberry S L, Dale E, Qin M, Ow D W
Plant Gene Expression Center, US Department of Agriculture, Albany, CA 94710.
Nucleic Acids Res. 1995 Feb 11;23(3):485-90. doi: 10.1093/nar/23.3.485.
Chromosomal rearrangements are useful genetic and breeding tools but are often difficult to detect and characterize. To more easily identify and define chromosome deletions and inversions, we have used the bacteriophage P1 Cre-lox site-specific recombination system to generate these events in plants. This involves three steps: (i) the introduction of two lox sites into one locus in a plant genome, including one site within a modified Ds transposon; (ii) Ac transposase-mediated transposition of the Ds-lox element to a new locus on the same chromosome; (iii) Cre-mediated site-specific recombination between the two lox sites that bracket a chromosome segment. We report the production of a deletion and three inversion events in tobacco. The utility of chromosomal segments bracketed by lox sites for targeted manipulation and cloning is discussed.
染色体重排是有用的遗传和育种工具，但往往难以检测和表征。为了更轻松地识别和定义染色体缺失和倒位，我们利用噬菌体P1 Cre-lox位点特异性重组系统在植物中产生这些事件。这包括三个步骤：（i）将两个lox位点引入植物基因组中的一个位点，包括在一个修饰的Ds转座子内的一个位点；（ii）Ac转座酶介导的Ds-lox元件转座到同一条染色体上的一个新位点；（iii）Cre介导的两个lox位点之间的位点特异性重组，这两个lox位点界定了一个染色体片段。我们报告了烟草中一个缺失和三个倒位事件的产生。讨论了由lox位点界定的染色体片段在靶向操作和克隆中的实用性。
