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    • 1. 发明申请
    • Cosmid Vector For Plant Transformation And Use Thereof
    • 用于植物转化和使用的粘粒载体
    • US20100132068A1
    • 2010-05-27
    • US12306163
    • 2007-06-25
    • Yoshimitsu TakakuraToshihiko KomariYuji IshidaToshiyuki KomoriYukoh HieiToshiki MineTeruyuki Imayama
    • Yoshimitsu TakakuraToshihiko KomariYuji IshidaToshiyuki KomoriYukoh HieiToshiki MineTeruyuki Imayama
    • A01H1/00C12N15/63
    • C12N15/82C12N15/8205
    • The present invention aims to provide novel vectors for plant transformation.The vectors of the present invention are cosmid vectors having a full length of 15 kb or less characterized in that: 1) they contain an origin of replication of an IncP plasmid, but do not contain any origin of replication of other plasmid groups; 2) they contain the trfA1 gene of an IncP plasmid; 3) they contain an oriT of an IncP plasmid; 4) they contain the incC1 gene of an IncP plasmid; 5) they contain a cos site of lambda phage and the cos site is located outside the T-DNA; 6) they contain a drug resistance gene expressed in E. coli and a bacterium of the genus Agrobacterium; 7) they contain a T-DNA right border sequence of a bacterium of the genus Agrobacterium; 8) they contain a T-DNA left border sequence of a bacterium of the genus Agrobacterium; 9) they contain a selectable marker gene for plant transformation located between 7) and 8) and expressed in a plant; and 10) they contain restriction endonuclease recognition site(s) located between 7) and 8) for cloning a foreign gene.
    • 本发明旨在提供用于植物转化的新型载体。 本发明的载体是全长为15kb或更小的粘粒载体,其特征在于:1)它们含有IncP质粒的复制起点,但不含有其他质粒组的任何复制起点; 2)它们含有IncP质粒的trfA1基因; 3)它们含有一个IncP质粒的oriT; 4)它们含有IncP质粒的incC1基因; 5)它们含有λ噬菌体的cos位点,cos位点位于T-DNA外部; 6)它们含有在大肠杆菌中表达的耐药基因和农杆菌属的细菌; 7)它们含有农杆菌属细菌的T-DNA右侧序列; 8)它们含有农杆菌属细菌的T-DNA左边界序列; 9)它们含有位于7)和8之间的植物转化的选择性标记基因并在植物中表达; 和10)它们含有位于7)和8之间的限制性内切核酸酶识别位点,用于克隆外来基因。
    • 2. 发明授权
    • Cosmid vector for plant transformation and use thereof
    • 用于植物转化的粘粒载体及其用途
    • US08298819B2
    • 2012-10-30
    • US12306163
    • 2007-06-25
    • Yoshimitsu TakakuraToshihiko KomariYuji IshidaToshiyuki KomoriYukoh HieiToshiki MineTeruyuki Imayama
    • Yoshimitsu TakakuraToshihiko KomariYuji IshidaToshiyuki KomoriYukoh HieiToshiki MineTeruyuki Imayama
    • C12N15/82C12N15/19C12N15/00
    • C12N15/82C12N15/8205
    • The present invention aims to provide novel vectors for plant transformation.The vectors of the present invention are cosmid vectors having a full length of 15 kb or less characterized in that: 1) they contain an origin of replication of an IncP plasmid, but do not contain any origin of replication of other plasmid groups; 2) they contain the trfA1 gene of an IncP plasmid; 3) they contain an oriT of an IncP plasmid; 4) they contain the incC1 gene of an IncP plasmid; 5) they contain a cos site of lambda phage and the cos site is located outside the T-DNA; 6) they contain a drug resistance gene expressed in E. coli and a bacterium of the genus Agrobacterium; 7) they contain a T-DNA right border sequence of a bacterium of the genus Agrobacterium; 8) they contain a T-DNA left border sequence of a bacterium of the genus Agrobacterium; 9) they contain a selectable marker gene for plant transformation located between 7) and 8) and expressed in a plant; and 10) they contain restriction endonuclease recognition site(s) located between 7) and 8) for cloning a foreign gene.
    • 本发明旨在提供用于植物转化的新型载体。 本发明的载体是全长为15kb或更小的粘粒载体,其特征在于:1)它们含有IncP质粒的复制起点,但不含有其他质粒组的任何复制起点; 2)它们含有IncP质粒的trfA1基因; 3)它们含有一个IncP质粒的oriT; 4)它们含有IncP质粒的incC1基因; 5)它们含有λ噬菌体的cos位点,cos位点位于T-DNA外部; 6)它们含有在大肠杆菌中表达的耐药基因和农杆菌属的细菌; 7)它们含有农杆菌属细菌的T-DNA右侧序列; 8)它们含有农杆菌属细菌的T-DNA左边界序列; 9)它们含有位于7)和8之间的植物转化的选择性标记基因并在植物中表达; 和10)它们含有位于7)和8之间的限制性内切核酸酶识别位点,用于克隆外来基因。
    • 6. 发明授权
    • Method of transforming monocotyledons using scutella of immature embryos
    • 使用未成熟胚的小檗转化单子叶植物的方法
    • US07939328B1
    • 2011-05-10
    • US08428238
    • 1994-09-01
    • Hideaki SaitoYuji IshidaYukoh HieiToshihiko Komari
    • Hideaki SaitoYuji IshidaYukoh HieiToshihiko Komari
    • C12N15/82
    • C12N15/8205
    • Disclosed is a method of transforming monocotyledons which necessitates only a short period from the transformation to the regeneration of a whole plant as compared with the conventional methods, thus reducing the frequency of occurrence of mutants, and can be generally applied to the plants for which any system of regenerating the whole plants from protoplasts has not been established, and in which the material to be used can be readily prepared without any particular apparatuses. The present invention provides a method for transforming monocotyledons comprising transforming scutellum of an immature embryo of a monocotyledon with a bacterium belonging to genus Agrobacterium containing a desired gene, which immature embryo has not been subjected to a dedifferentiation treatment, to obtain a transformant.
    • 本发明公开了一种转化单子叶植物的方法,其与常规方法相比仅需要从整个植物的转化到再生的短的时间,从而降低突变体的发生频率,并且通常可以应用于任何 尚未建立从原生质体再生整株植物的系统,其中待使用的材料可以在没有任何特定装置的情况下容易地制备。 本发明提供一种用于将单子叶植物的未成熟胚的鳞片状细菌与含有未成熟胚的未分化处理的所需基因的农杆菌属的农杆菌属转化成单转基因的方法,得到转化体。
    • 7. 发明授权
    • Method for introducing two T-DNAS into plants and vectors therefor
    • 将两种T-DNAS引入植物及其载体的方法
    • US5731179A
    • 1998-03-24
    • US500952
    • 1995-08-08
    • Toshihiko KomariYasuhito SaitoYukoh Hiei
    • Toshihiko KomariYasuhito SaitoYukoh Hiei
    • C12N15/74C12N15/82C12N15/84C12N15/05A01H5/00C12N1/20
    • C12N15/743C12N15/8205
    • The invention provides a method for transforming a plant through a bacterium belonging to genus Agrobacterium, comprising transforming plant cells simultaneously with a first T-DNA (1) and a second T-DNA (2); and selecting the cells which acquired drug resistance; the first T-DNA (1) containing a gene giving the drug resistance, which functions in the plant; the second T-DNA (2) containing a desired DNA fragment to be introduced into the plant, the second T-DNA (2) being contained in a hybrid vector; the hybrid vector being prepared by homologous recombination between an acceptor vector and an intermediate vector in the bacterium belonging to genus Agrobacterium; the acceptor vector containing at least (a) a DNA region having a function to replicate a plasmid in the bacterium belonging to genus Agrobacterium and Escherichia coli, (b) a DNA region containing virB gene and virG gene in virulence region of Ti plasmid pTiBo542 of Agrobacterium tumefaciens, and (c) a DNA region which is homologous with a part of the intermediate vector, which is subjected to homologous recombination in the bacterium belonging to genus Agrobacterium; the intermediate vector containing at least (i) a DNA region having a function to replicate a plasmid in Escherichia coli, which does not function in the bacterium belonging to genus Agrobacterium, (ii) a DNA region which is homologous with a part of the acceptor vector, which is subjected to homologous recombination in the bacterium belonging to genus Agrobacterium, and (iii) a DNA region which constitutes at least a part of the second T-DNA.
    • PCT No.PCT / JP94 / 02049 Sec。 371日期:1995年8月8日 102(e)日期1995年8月8日PCT 1994年12月6日PCT PCT。 公开号WO95 / 16031 日期:1995年6月15日本发明提供了通过农杆菌属细菌转化植物的方法,其包括与第一T-DNA(1)和第二T-DNA(2)同时转化植物细胞; 并选择获得耐药性的细胞; 第一个含有赋予药物抗性的基因的T-DNA(1)在植物中起作用; 第二T-DNA(2)含有待引入植物的所需DNA片段,第二T-DNA(2)包含在杂交载体中; 杂交载体通过在土壤杆菌属的细菌中的受体载体和中间载体之间的同源重组而制备; 所述受体载体至少含有(a)具有复制属于农杆菌属和大肠杆菌属的细菌中的质粒的功能的DNA区域,(b)含有TiBp质粒pTiBo542的毒力区域中的virB基因和virG基因的DNA区域 根癌农杆菌,和(c)与属于农杆菌属细菌的同源重组的中间载体的一部分同源的DNA区域; 所述中间载体至少含有(i)具有在大肠杆菌中复制质粒的功能的DNA区域,其在属于农杆菌属的细菌中不起作用,(ii)与受体的一部分同源的DNA区域 载体,其在属于农杆菌属的细菌中进行同源重组,和(iii)构成第二T-DNA的至少一部分的DNA区。
    • 8. 发明授权
    • Method for transforming monocotyledons
    • 单子叶植物的转化方法
    • US07060876B2
    • 2006-06-13
    • US09229324
    • 1999-01-13
    • Yukoh HieiToshihiko Komari
    • Yukoh HieiToshihiko Komari
    • A01H1/00A01H5/00C07H2/04C12N15/82C12N15/84
    • C12N15/8205
    • The invention relates to a method for transforming a monocotyledonous plant. The time required from transformation to regeneration of a plant is shorter using the inventive method so that the frequency of emergence of mutants is smaller than the conventional methods. The inventive method may be generally applied even to the plants for which a regeneration method from a protoplast to a plant has not been established, and with which the preparation of the material to be subjected to the method is easy. That is, the present invention provides a method for transforming a monocotyledonous plant, comprising contacting a cultured tissue of said monocotyledonous plant during dedifferentiation thereof obtained by culturing an explant on a dedifferentiation-inducing medium for less than 7 days with a bacterium belonging to the genus Agrobacterium containing a super binary vector having the virulence region of a Ti plasmid, left and right border sequences of T-DNA of a Ti plasmid or an Ri plasmid of a bacterium belonging to the genus Agrobacterium, and a desired gene located between said left and right border sequences.
    • 本发明涉及一种用于转化单子叶植物的方法。 使用本发明的方法从植物的转化到再生所需的时间更短,使得突变体的出现频率比常规方法小。 本发明的方法通常可以应用于从原生质体到植物的再生方法尚未建立的植物,并且通过该方法对待处理的材料的制备是容易的。 也就是说,本发明提供了一种用于转化单子叶植物的方法,包括使所述单子叶植物的去分化过程中的培养的组织与通过在去分化诱导培养基上培养少于7天的外植体与属于该属的细菌接触 含有具有Ti质粒的毒力区域的超级二元载体的农杆菌,Ti质粒的T-DNA的左侧和右侧边界序列或属于农杆菌属的细菌的Ri质粒,以及位于所述左侧和 右边界序列
    • 9. 发明授权
    • Vectors for transforming plants
    • 用于转化植物的载体
    • US07087812B1
    • 2006-08-08
    • US09856976
    • 1999-09-30
    • Yoshiki KurayaToshihiko KomariYukoh Hiei
    • Yoshiki KurayaToshihiko KomariYukoh Hiei
    • A01H1/00C12N15/82C12N1/20C07H2/04
    • C12N15/8205
    • Vectors for transforming plants with the use of agrobacteria which have been modified so as to elevate the possibility of the recognition of the border sequences of the vectors by vir proteins of the agrobacteria, thereby lowering the possibility of the transfer of DNAs other than T-DNA into plant chromosomes. More particularly, the above-vectors are those to be used in transforming plants which have right and left border sequences which can be recognized by the vir proteins of the agrobacteria, a T-DNA sequence which is located between these border sequences and into which a gene to be transferred into plants can be inserted, and a replication origin enabling the replication of the vectors in bacteria, characterized by having a plural number of left border sequences.
    • 用于使用农杆菌转化植物的载体,其被修饰以提高通过农杆菌的vir蛋白识别载体的边界序列的可能性,从而降低T-DNA以外的DNA的转移的可能性 进入植物染色体。 更具体地说,上述载体是用于转化具有可被农杆菌的vir蛋白识别的右边界和左侧边界序列的转化植物,位于这些边界序列之间的T-DNA序列,其中 可以插入待转移到植物中的基因,以及能够复制细菌中的载体的复制起点,其特征在于具有多个左边界序列。
    • 10. 发明授权
    • Method for transforming monocotyledons
    • 单子叶植物的转化方法
    • US5591616A
    • 1997-01-07
    • US193058
    • 1994-05-03
    • Yokoh HieiToshihiko Komari
    • Yokoh HieiToshihiko Komari
    • A01H1/00C12N5/10C12N15/82C12M15/00C12R1/41
    • C12N15/8205
    • A method for transforming a monocotyledon by which the time required from transformation to regeneration of a plant is shorter so that the frequency of emergence of mutants is smaller than the conventional methods, which may be generally applied even to the plants for which the regeneration method from a protoplast to a plant has not been established, and with which the preparation of the material to be subjected to the method is easy. That is, the present invention provides a method for transforming a monocotyledon comprising transforming a cultured tissue during dedifferentiation process or a dedifferentiated cultured tissue of said monocotyledon with a bacterium belonging to genus Agrobacterium containing a desired gene.
    • PCT No.PCT / JP93 / 00925 Sec。 371日期:1994年5月3日 102(e)日期1994年5月3日PCT提交1993年7月6日PCT公布。 第WO94 / 00977号公报 日期1994年1月20日一种用于转化单子叶植物的方法,其中从转化到植物再生所需的时间更短,使得突变体的出现频率小于常规方法,甚至通常适用于植物 从原生质体到植物的再生方法尚未建立,用于该方法的材料的制备容易。 也就是说,本发明提供了一种用于转化单子叶植物的方法,包括在去分化过程中转化培养的组织或将所述单子叶植入物的去分化培养的组织与含有所需基因的农杆菌属的细菌进行转化。