Bayley C C, Morgan M, Dale E C, Ow D W
Plant Gene Expression Center, USDA/ARS, Albany, CA 94710.
Plant Mol Biol. 1992 Jan;18(2):353-61. doi: 10.1007/BF00034962.
The Cre-lox site-specific recombination system of bacteriophage P1 was used to excise a firefly luciferase (luc) gene which had previously been incorporated into the tobacco genome. The excision event was due to site-specific DNA recombination between two lox sequences flanking the luc gene and was catalyzed by the Cre recombinase introduced by cross-fertilization. Recombination resulted in the fusion of a promoter with a distally located hygromycin phosphotransferase (hpt) coding sequence and the excision event was monitored as a phenotypic change from expression of luc to expression of hpt. The efficiency of recombination was estimated from the exchange of gene activity and confirmed by molecular analysis. The relevance to potential applications of site-specific deletion-fusion events for chromosome engineering are discussed.
利用噬菌体P1的Cre-lox位点特异性重组系统切除先前已整合到烟草基因组中的萤火虫荧光素酶(luc)基因。切除事件是由于luc基因两侧的两个lox序列之间发生位点特异性DNA重组,并由通过杂交引入的Cre重组酶催化。重组导致一个启动子与位于远端的潮霉素磷酸转移酶(hpt)编码序列融合,并且切除事件通过从luc表达转变为hpt表达的表型变化进行监测。重组效率通过基因活性的交换来估计,并通过分子分析得到证实。文中还讨论了位点特异性缺失-融合事件在染色体工程潜在应用中的相关性。