Terada Rie, Urawa Hiroko, Inagaki Yoshishige, Tsugane Kazuo, Iida Shigeru
National Institute for Basic Biology, Okazaki, 444-8585, Japan.
Nat Biotechnol. 2002 Oct;20(10):1030-4. doi: 10.1038/nbt737. Epub 2002 Sep 9.
Modification of genes through homologous recombination, termed gene targeting, is the most direct method to characterize gene function. In higher plants, however, the method is far from a common practice. Here we describe an efficient and reproducible procedure with a strong positive/negative selection for gene targeting in rice, which feeds more than half of the world's population and is an important model plant. About 1% of selected calli and their regenerated fertile plants were heterozygous at the targeted locus, and only one copy of the selective marker used was found at the targeted site in their genomes. The procedure's applicability to other genes will make it feasible to obtain various gene-targeted lines of rice.
通过同源重组对基因进行修饰，即基因打靶，是表征基因功能的最直接方法。然而，在高等植物中，该方法远未成为一种常规做法。在此，我们描述了一种高效且可重复的程序，该程序对水稻基因打靶具有强大的正/负选择作用，水稻养活了世界一半以上的人口，是一种重要的模式植物。约1%的所选愈伤组织及其再生的可育植株在靶位点处为杂合子，并且在其基因组的靶位点仅发现了所使用的一个选择性标记拷贝。该程序对其他基因的适用性将使获得各种水稻基因打靶品系成为可能。
