首页> 外国专利> METHOD FOR INTEGRATING OF EXZOGENOUS DNA INTO GENOME OF A PLANT CELL AND METHOD FOR PRODUCING FERTILE TRANSGENIC PLANT INTEGRATED INTO THE GENOME OF THE EXZOGENOUS DNA

METHOD FOR INTEGRATING OF EXZOGENOUS DNA INTO GENOME OF A PLANT CELL AND METHOD FOR PRODUCING FERTILE TRANSGENIC PLANT INTEGRATED INTO THE GENOME OF THE EXZOGENOUS DNA

机译:将外源DNA整合到植物细胞基因组中的方法和生产整合到外源DNA基因组中的铁转基因植物的方法

摘要

1. A method for integrating of a exzogenous DNA into the genome of a plant cell wherein comprising transforming said cell of said DNA with a DNA fragment consisting of the DNA of interest bounded by T-DNA border sequences; and with at least one chimeric DNA or RNA molecule capable of expressing in said plant cell one or more proteins mediating the integration of the exzogenous DNA, wherein these proteins are encoded by the virulence region of an Agrobacterium Ti or Ri plasmid so that said proteins are present in a plant cell in a sufficient amount after introducing the exzogenous DNA into said plant cell but before the integration of the exzogenous DNA into the genome of said plant cell. 2. The method of claim 1 wherein the exzogenous DNA and chimeric DNA or RNA are introduced simultaneously. 3. The method of claim 1 wherein one or more proteins mediating the integration of the exzogenous DNA encode VirD1, VirD2, VirC1, VirC2 or VirE2. 4. The method of claim 3 wherein one or more proteins mediating the integration of the exzogenous DNA encode VirD1 and VirD2, or a combination of VirD1, VirD2 and VirE2. 5. The method of claim 3 wherein one or more proteins mediating the integration of the exzogenous DNA encode VirD2. 6. The method of claim 1 wherein the exzogenous DNA bounded by T-DNA border sequences is part of a plant viral vector. 7. The method of claim 1 wherein said DNA gene is part of a plant viral vector. 8. The method of claim 1 wherein the exzogenous DNA bounded by T-DNA border sequences and said DNA gene are part of a plant viral vector. 9. The method of claim 1herein said plant cell is a cell of a dicotyledonous plant. 10. The method of claim 9 wherein said dicotyledonous plant is selected from th e group consisting of tobacco, cotton, oilseed rape, and soybean. 11. The method of claim 1 wherein said plant cell is a cell of a monocotyledonous plant. 12. The method of claim 13 wherein said monocotyledonous plant is selected from the group consisting of maize, wheat and rice. 13. The method of claim 1 wherein said exzogenous DNA comprises a chimeric gene. 14. The method of claim 15 wherein said chimeric gene is capable of expressing a protein in said plant cell. 15. The method of claim 1 wherein said exzogenous DNA bounded by T-DNA border sequences and said DNA gene are components of a single DNA molecule. 16. A method for producing a fertile transgenic plant having DNA bounded by T-DN A border sequences integrated into the genome comprising transforming said DNA into the genome of a plant cell according to the method of claim 3; and regenerating the transformed plant cell to a fertile transgenic plant. 17. The method of claim 16 wherein said fertile transgenic plant is selected from the group consisting of tobacco, cotton, oilseed rape, soybean, maize, wheat and rice. 18. The method of claim 16 wherein said fertile transgenic plant is selected from the group consisting of maize, wheat, rye, sorghum, rice, barley, millet, sod-forming and forage grasses and also sugar cane sugar beet, oilseed rape, banana, poplar nut, tobacco and soybean.
机译:1.一种将外源DNA整合到植物细胞基因组中的方法,其中包括用由T-DNA边界序列所结合的目的DNA组成的DNA片段转化所述DNA的所述细胞;并且具有至少一种能够在所述植物细胞中表达一种或多种介导外源DNA整合的蛋白质的嵌合DNA或RNA分子,其中这些蛋白质由农杆菌Ti或Ri质粒的毒力区编码,使得所述蛋白质为在将外源DNA引入所述植物细胞之后但在将外源DNA整合到所述植物细胞的基因组中之前,存在于植物细胞中的RNA的量足够。 2.权利要求1的方法,其中外源DNA和嵌合DNA或RNA被同时引入。 3.权利要求1的方法,其中介导外源DNA整合的一种或多种蛋白质编码VirD1,VirD2,VirC1,VirC2或VirE2。 4.权利要求3的方法,其中介导外源DNA整合的一种或多种蛋白质编码VirD1和VirD2,或VirD1,VirD2和VirE2的组合。 5.权利要求3的方法,其中一种或多种介导外源DNA整合的蛋白质编码VirD2。 6.权利要求1的方法,其中由T-DNA边界序列结合的外源DNA是植物病毒载体的一部分。 7.权利要求1的方法,其中所述DNA基因是植物病毒载体的一部分。 8.权利要求1的方法,其中由T-DNA边界序列结合的外源DNA和所述DNA基因是植物病毒载体的一部分。 9.权利要求1的方法,其中所述植物细胞是双子叶植物的细胞。 10.权利要求9的方法,其中所述双子叶植物选自烟草,棉花,油菜和大豆。 11.权利要求1的方法,其中所述植物细胞是单子叶植物的细胞。 12.权利要求13的方法,其中所述单子叶植物选自玉米,小麦和水稻。 13.权利要求1的方法,其中所述外源DNA包含嵌合基因。 14.权利要求15的方法,其中所述嵌合基因能够在所述植物细胞中表达蛋白质。 15.权利要求1的方法,其中所述由T-DNA边界序列结合的外源DNA和所述DNA基因是单个DNA分子的组分。 16.一种产生具有被T-DN A边界序列结合的DNA的可育转基因植物的方法,该DNA被整合到基因组中,其包括根据权利要求3所述的方法将所述DNA转化成植物细胞的基因组。并将转化的植物细胞再生为能育的转基因植物。 17.权利要求16的方法,其中所述可育转基因植物选自烟草,棉花,油菜,大豆,玉米,小麦和大米。 18.权利要求16的方法,其中所述可育转基因植物选自玉米,小麦,黑麦,高粱,水稻,大麦,小米,成草草和饲料草,以及甘蔗甜菜,油菜,香蕉,白杨仁,烟草和大豆。

著录项

  • 公开/公告号EA001039B1

    专利类型

  • 公开/公告日2000-08-28

    原文格式PDF

  • 申请/专利权人 NOVARTIS AG;

    申请/专利号EA19980000324

  • 发明设计人 CHILTON MARY-DELL;HANSEN GENEVIEVE;

    申请日1996-09-12

  • 分类号C12N15/82;C12N5/10;A01H5/00;

  • 国家 EA

  • 入库时间 2022-08-22 01:56:38

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