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公开(公告)号：US07223601B2
公开(公告)日：2007-05-29
申请号：US10353445
申请日：2003-01-29
申请人： Christopher L. Baszczynski , Leszek Alexander Lyznik , William J. Gordon-Kamm , Xueni Guan , Aragula Gururaj Rao , Laura A. Tagliani
发明人： Christopher L. Baszczynski , Leszek Alexander Lyznik , William J. Gordon-Kamm , Xueni Guan , Aragula Gururaj Rao , Laura A. Tagliani
IPC分类号： C12N15/87
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，所述方法和组合物涉及使用两个位点特异性重组酶的靶位点的组合和具有双重靶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。
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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。
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公开(公告)号：US06555673B1
公开(公告)日：2003-04-29
申请号：US09556163
申请日：2000-04-21
IPC分类号： C07H2104
CPC分类号： C12N9/0008 , C12N15/8216 , C12N15/8222 , C12N15/8225 , C12N15/8226 , C12N15/8279
摘要： Synthetic elements for enhancing expression of genes in plant cells are disclosed. These include a promoter with a “TATA to start” sequence containing 64% or greater GC content and an synthetic upstream element incorporating several OCS binding motifs and novel flanking sequences. Upstream activating regions (UARs) are also disclosed that can further increase the constitutive transcriptional activity when they are operably linked to said promoter and/or the synthetic upstream element. In particular, the nucleotide sequence of the UAR of the maize Ubi-1 gene is provided and its use in expression cassettes and vectors containing these promoter elements. Cells and plants transformed with these vectors are further provided. These include a transgenic sunflower expressing an exogenous oxalate oxidase gene at a high level under the transcriptional control of a recombinant promoter having at least one upstream activating region of the 35S CaMV promoter.
摘要翻译： 公开了用于增强植物细胞中基因表达的合成元件。 这些包括具有含有64％或更高GC含量的“TATA起始”序列的启动子和包含若干OCS结合基序和新侧翼序列的合成上游元件。 还公开了上游活化区（UAR），当它们可操作地连接到所述启动子和/或合成上游元件时，可以进一步增加组成型转录活性。 特别地，提供了玉米Ubi-1基因的UAR的核苷酸序列，并且其用于表达盒和含有这些启动子元件的载体。 进一步提供用这些载体转化的细胞和植物。 这些包括在具有35S CaMV启动子的至少一个上游活化区的重组启动子的转录控制下在高水平表达外源性草酸氧化酶基因的转基因向日葵。
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公开(公告)号：US6072050A
公开(公告)日：2000-06-06
申请号：US28819
申请日：1998-02-24
CPC分类号： C12N9/0008 , C12N15/8216 , C12N15/8222 , C12N15/8225 , C12N15/8226 , C12N15/8279
摘要： Synthetic elements for enhancing expression of genes in plant cells are disclosed. These include a promoter with a "TATA to start" sequence containing 64% or greater GC content and an synthetic upstream element incorporating several OCS binding motifs and novel flanking sequences. Upstream activating regions (UARs) are also disclosed that can further increase the constitutive transcriptional activity when they are operably linked to said promoter and/or the synthetic upstream element. In particular, the nucleotide sequence of the UAR of the maize Ubi-1 gene is provided and its use in expression cassettes and vectors containing these promoter elements. Cells and plants transformed with these vectors are further provided. These include a transgenic sunflower expressing an exogenous oxalate oxidase gene at a high level under the transcriptional control of a recombinant promoter having at least one upstream activating region of the 35S CaMV promoter.
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公开(公告)号：US06369298B1
公开(公告)日：2002-04-09
申请号：US09056418
申请日：1998-04-07
申请人： Tishu Cai , Dorothy A. Pierce , Laura A. Tagliani , Zuo-Yu Zhao
发明人： Tishu Cai , Dorothy A. Pierce , Laura A. Tagliani , Zuo-Yu Zhao
IPC分类号： A01H100
CPC分类号： C12N15/8205
摘要： Methods and compositions for the efficient transformation of sorghum is provided. The method involves infection with Agrobacterium, particularly those comprising a super-binary vector. In this manner, any gene of interest can be introduced into the sorghum plant. The transformed gene will be flanked by at least one T-DNA border and present in the transformed sorghum in low copy number. Transformed sorghum, cells, tissues, plants, and seed are also provided. The invention encompasses regenerated, fertile sorghum plants, transgenic seeds produced therefrom, T1 and subsequent generations.
摘要翻译： 提供了高效转化高粱的方法和组成。 该方法涉及用农杆菌感染，特别是包含超二元载体的那些。 以这种方式，可以将任何感兴趣的基因引入高粱植物中。 转化的基因将侧接至少一个T-DNA边界并以低拷贝数存在于转化的高粱中。 还提供了转化的高粱，细胞，组织，植物和种子。 本发明包括再生的，可育的高粱植物，由其产生的转基因种子，T1和后代。
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公开(公告)号：US5976806A
公开(公告)日：1999-11-02
申请号：US85902
申请日：1998-05-27
CPC分类号： C12Q1/25 , C12Q1/6897 , G01N2333/9015
摘要： A quantitative and functional DNA ligase assay is disclosed. The assay involves the disruption, using restriction enzymes, of a plasmid containing a reporter gene, followed by a ligation step performed in the presence of the biological sample being assayed for DNA ligase activity. The ligation reaction products are then subjected to a coupled transcription-translation reaction, and the extent of DNA ligation is then quantified.
摘要翻译： 公开了定量和功能性DNA连接酶测定。 该测定涉及使用限制酶破坏含有报道基因的质粒，随后在待测DNA生物样品的DNA连接酶活性的存在下进行连接步骤。 然后将连接反应产物进行偶联转录翻译反应，然后定量DNA连接的程度。
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