Lyznik L A, Mitchell J C, Hirayama L, Hodges T K
Department of Botany and Plant Pathology, Purdue University, West Lafayette, IA 47907.
Nucleic Acids Res. 1993 Feb 25;21(4):969-75. doi: 10.1093/nar/21.4.969.
We have demonstrated that a yeast FLP/FRT site-specific recombination system functions in maize and rice protoplasts. FLP recombinase activity was monitored by reactivation of beta-glucuronidase (GUS) expression from vectors containing the gusA gene inactivated by insertion of two FRTs (FLP recombination targets) and a 1.31 kb DNA fragment. The stimulation of GUS activity in protoplasts cotransformed with vectors containing FRT inactivated gusA gene and a chimeric FLP gene depended on both the expression of the FLP recombinase and the presence and structure of the FRT sites. The FLP enzyme could mediate inter- and intramolecular recombination in plant protoplasts. These results provide evidence that a yeast recombination system can function efficiently in plant cells, and that its performance can be manipulated by structural modification of the FRT sites.
我们已经证明酵母FLP/FRT位点特异性重组系统在玉米和水稻原生质体中发挥作用。通过重新激活β-葡萄糖醛酸酶（GUS）的表达来监测FLP重组酶的活性，该表达来自含有gusA基因的载体，该基因因插入两个FRT（FLP重组靶标）和一个1.31 kb DNA片段而失活。在用含有FRT失活gusA基因的载体和嵌合FLP基因共转化的原生质体中，GUS活性的刺激取决于FLP重组酶的表达以及FRT位点的存在和结构。FLP酶可以介导植物原生质体中的分子间和分子内重组。这些结果提供了证据，表明酵母重组系统可以在植物细胞中高效发挥作用，并且其性能可以通过FRT位点的结构修饰来操纵。
