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    Method of homologous recombination followed by in vivo selection of DNA amplification
    4.
    发明授权
    Method of homologous recombination followed by in vivo selection of DNA amplification 失效
    同源重组方法，然后进行DNA扩增的体内选择

    公开(公告)号：US5925544A

    公开(公告)日：1999-07-20

    申请号：US954391

    申请日：1997-10-20

    申请人： Steen Troels J.o slashed.rgensen

    发明人： Steen Troels J.o slashed.rgensen

    IPC分类号： C12N15/64 ,  C12N15/90 ,  C12P21/00 ,  C07H21/04 ,  C12P19/34 ,  C12Q1/68

    CPC分类号： C12N15/64 ,  C12N15/902

    摘要： A method of amplifying in vivo a DNA sequence B present in a genome of a parent cell, comprising (a) integrating in a genome of said cell a DNA construct comprising the structure C-M-A-D, in which both A and C denote a DNA sequence which is homologous with a genomic DNA fragment either flanking or overlapping the DNA sequence B, the sequence C being located in the opposite end the sequence B as compared to A, D denotes a DNA sequence which is homologous with a genomic DNA fragment located distal for C as compared to B, and M denotes a DNA sequence encoding a selection marker, (b) selecting for cells in which the DNA sequence M has been integrated in the genome, which cells comprise, in any orientation, the structure A-B-C-M-A-D and (c) propagating the cells selected in step (b) under increasing selection pressure to obtain a cell which has obtained an increased number of genomically integrated copies of the DNA sequences B and M.

    摘要翻译： 在体内扩增存在于亲本细胞基因组中的DNA序列B的方法，包括（a）在所述细胞的基因组中整合含有结构CMAD的DNA构建体，其中A和C都表示DNA序列， 与DNA序列B侧翼或重叠的基因组DNA片段同源，与A相比，序列C位于序列B的相对端，D表示与位于C末端的基因组DNA片段同源的DNA序列 与B相比，M表示编码选择标记的DNA序列，（b）选择其中DNA序列M已经整合到基因组中的细胞，该细胞以任何方向包含结构ABCMAD和（c）传播的结构 在增加的选择压力下在步骤（b）中选择的细胞，以获得获得DNA序列B和M的增加的基因整合拷贝数的细胞。

    DNA integration by transposition
    5.
    发明授权
    DNA integration by transposition 失效
    通过转座进行DNA整合

    公开(公告)号：US5882888A

    公开(公告)日：1999-03-16

    申请号：US875154

    申请日：1997-07-17

    申请人： Steen Troels J.o slashed.rgensen

    发明人： Steen Troels J.o slashed.rgensen

    IPC分类号： C12N15/09 ,  C12N1/21 ,  C12N15/75 ,  C12N15/90 ,  C12P21/02 ,  C12R1/07 ,  C12R1/225 ,  C12P21/00 ,  C07H21/04 ,  C12N15/70

    CPC分类号： C12N15/75 ,  C12N15/90

    摘要： Multicopy strains of gram-positive bacteria carrying multiple copies of a DNA sequence of interest may be constructed by use of a method involving introduction of a DNA construct comprising the DNA sequence of interest into the genome of the recipient cell by transposition and subsequent deletion of a marker gene used for selection of the cells having received the DNA construct by a resolution system. The multicopy strains are preferably free from a gene encoding an undesirable marker such as an antibiotic resistance marker.

    摘要翻译： PCT No.PCT / DK96 / 00038 Sec。 371日期1997年7月17日 102（e）日期1997年7月17日PCT 1996年1月23日PCT PCT。 公开号WO96 / 23073 日期1996年8月1日携带多个目的DNA序列拷贝的革兰氏阳性细菌的多拷贝菌株可以通过使用包括通过转座将包含感兴趣的DNA序列的DNA构建体引入受体细胞的基因组的方法来构建 以及随后删除用于通过分辨系统选择接受DNA构建体的细胞的标记基因。 多拷贝菌株优选不含编码不良标志物如抗生素抗性标记物的基因。

    Amplification of genomic DNA by site specific integration of a selectable marker construct
    6.
    发明授权
    Amplification of genomic DNA by site specific integration of a selectable marker construct 失效
    通过选择性标记构建体的位点特异性整合扩增基因组DNA

    公开(公告)号：US5733753A

    公开(公告)日：1998-03-31

    申请号：US751282

    申请日：1996-11-18

    申请人： Steen Troels J.o slashed.rgensen

    发明人： Steen Troels J.o slashed.rgensen

    IPC分类号： C12N15/64 ,  C12N15/67 ,  C12N15/90 ,  C12P19/34 ,  C07H21/02 ,  C12Q1/68

    CPC分类号： C12N15/67 ,  C12N15/64 ,  C12N15/902

    摘要： A method of amplifying in vivo a DNA sequence B present in a genome of a parent cell, comprising (a) integrating in a genome of said cell a DNA construct comprising the structure C-M-A-D, in which both A and C denote a DNA sequence which is homologous with a genomic DNA fragment either flanking or overlapping the DNA sequence B, the sequence C being located in the opposite end the sequence B as compared to A, D denotes a DNA sequence which is homologous with a genomic DNA fragment located distal for C as compared to B, and M denotes a DNA sequence encoding a selection marker, (b) selecting for cells in which the DNA sequence M has been integrated in the genome, which cells comprise, in any orientation, the structure A-B-C-M-A-D and (c) propagating the cells selected in step (b) under increasing selection pressure to obtain a cell which has obtained an increased number of genomically integrated copies of the DNA sequences B and M.

    摘要翻译： 在体内扩增存在于亲本细胞基因组中的DNA序列B的方法，包括（a）在所述细胞的基因组中整合含有结构CMAD的DNA构建体，其中A和C都表示DNA序列， 与DNA序列B侧翼或重叠的基因组DNA片段同源，与A相比，序列C位于序列B的相对端，D表示与位于C末端的基因组DNA片段同源的DNA序列 与B相比，M表示编码选择标记的DNA序列，（b）选择其中DNA序列M已经整合到基因组中的细胞，该细胞以任何方向包含结构ABCMAD和（c）传播的结构 在增加的选择压力下在步骤（b）中选择的细胞，以获得获得DNA序列B和M的增加的基因整合拷贝数的细胞。