公开(公告)号：US06262341B1

公开(公告)日：2001-07-17

申请号：US09193503

申请日：1998-11-17

申请人： Christopher L. Baszczynski ,  Leszek Alexander Lyznik ,  William J. Gordon-Kamm ,  Xueni Guan ,  Argula Gururaj Rao ,  Laura A. Tagliani

发明人： Christopher L. Baszczynski ,  Leszek Alexander Lyznik ,  William J. Gordon-Kamm ,  Xueni Guan ,  Argula Gururaj Rao ,  Laura A. Tagliani

IPC分类号： C12N504

CPC分类号： C12N15/90 ,  C07K2319/00 ,  C12N9/00 ,  C12N15/8203 ,  C12N15/8205 ,  C12N15/8213

摘要： Compositions and methods for introducing a DNA of interest into a genomic target site are provided. In particular, the methods and compositions involve the use of a combination of target sites for two site specific recombinases and expression of a chimeric recombinase with dual target site specificity. Thus, the compositions comprise novel site-specific recombinases with specificities to multiple target sites, and nucleotide sequences and expression cassettes encoding these recombinases or target sites. The methods involve transforming a eukaryotic cell having target sites for the novel recombinase with a DNA of interest that is flanked by corresponding target sites. Expression of the recombinase results in integration of the DNA of interest into the genome of the cell. The compositions and methods of the invention have use in the construction of stably transformed eukaryotic cells, and in particular, plant cells. The methods result in the efficient targeted genomic integration of DNA by site-specific recombination.

摘要翻译： 提供了将目的DNA引入基因组靶位点的组合物和方法。 特别地，所述方法和组合物包括使用两个位点特异性重组酶的靶位点的组合和具有双靶点位点特异性的嵌合重组酶的表达。 因此，组合物包含对多个靶位点具有特异性的新的位点特异性重组酶，以及编码这些重组酶或靶位点的核苷酸序列和表达盒。 所述方法包括用具有相应靶位点侧翼的感兴趣的DNA转化具有新重组酶靶位点的真核细胞。 重组酶的表达导致感兴趣的DNA整合到细胞的基因组中。 本发明的组合物和方法用于构建稳定转化的真核细胞，特别是植物细胞。 该方法通过位点特异性重组导致DNA的有效靶向基因组整合。