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    Method for Identifying Drug-Sensitizing Antisense DNA Fragments and Use Thereof
    1.
    发明申请
    Method for Identifying Drug-Sensitizing Antisense DNA Fragments and Use Thereof 审中-公开
    鉴定药物敏感反义DNA片段及其用途的方法

    公开(公告)号：US20110143359A1

    公开(公告)日：2011-06-16

    申请号：US13033128

    申请日：2011-02-23

    申请人： Robert Haselbeck ,  Mark Hilgers ,  Karen Shaw ,  Vickie Brown-Driver ,  Kedar GC ,  John M. Finn ,  Mark Stidham

    发明人： Robert Haselbeck ,  Mark Hilgers ,  Karen Shaw ,  Vickie Brown-Driver ,  Kedar GC ,  John M. Finn ,  Mark Stidham

    IPC分类号： C12Q1/68

    CPC分类号： C12N15/111 ,  C12N15/1137 ,  C12N2310/11 ,  C12N2320/11 ,  C12N2330/30 ,  C12Y101/01158 ,  C12Y105/01003 ,  C12Y205/01031 ,  C12Y601/0101

    摘要： The invention provides a method for generating and selecting drug-sensitizing antisense DNA fragments. In one embodiment, the method includes identifying a gene of interest using knowledge of bacterial physiology, biochemistry, genetics, genomics, and other means. The method includes PCR amplification of a gene of interest using genomic DNA as a template; fragmentation of the DNA by sonication or other means; selecting DNA fragments no longer than 400 base pairs; ligating the DNA fragments into a suitable expression plasmid with a selectable marker; transforming the plasmids containing the DNA fragments into the organism from which the gene of interest originated; and selecting clones from transformed cells that show a phenotypic difference of the clone grown in the presence of the inducer relative to the phenotype in the absence of inducer.

    摘要翻译： 本发明提供了生产和选择药物敏化反义DNA片段的方法。 在一个实施方案中，该方法包括使用细菌生理学，生物化学，遗传学，基因组学和其他方法的知识来鉴定感兴趣的基因。 该方法包括使用基因组DNA作为模板对感兴趣的基因进行PCR扩增; 通过超声处理或其他方法破碎DNA; 选择不超过400个碱基对的DNA片段; 将DNA片段连接到具有选择性标记的合适的表达质粒中; 将含有DNA片段的质粒转化为目标基因来源的生物体; 并选择来自转化细胞的克隆，其在不存在诱导物的情况下显示在诱导物存在下生长的克隆相对于表型的表型差异。

    Method for identifying drug-sensitizing antisense DNA fragments and use thereof
    2.
    发明授权
    Method for identifying drug-sensitizing antisense DNA fragments and use thereof 有权
    用于鉴定药物敏化反义DNA片段的方法及其用途

    公开(公告)号：US07910337B2

    公开(公告)日：2011-03-22

    申请号：US11636394

    申请日：2006-12-08

    申请人： Robert Haselbeck ,  Mark Hilgers ,  Karen Shaw ,  Vickie Brown-Driver ,  Kedar Gc ,  John M. Finn ,  Mark Stidham

    发明人： Robert Haselbeck ,  Mark Hilgers ,  Karen Shaw ,  Vickie Brown-Driver ,  Kedar Gc ,  John M. Finn ,  Mark Stidham

    IPC分类号： C12P19/34 ,  C07H21/04 ,  C12N15/63 ,  C07K16/00

    CPC分类号： C12N15/111 ,  C12N15/1137 ,  C12N2310/11 ,  C12N2320/11 ,  C12N2330/30 ,  C12Y101/01158 ,  C12Y105/01003 ,  C12Y205/01031 ,  C12Y601/0101

    摘要： The invention provides a method for generating and selecting drug-sensitizing antisense DNA fragments. In one embodiment, the method includes identifying a gene of interest using knowledge of bacterial physiology, biochemistry, genetics, genomics, and other means. The method includes PCR amplification of a gene of interest using genomic DNA as a template; fragmentation of the DNA by sonication or other means; selecting DNA fragments no longer than 400 base pairs; ligating the DNA fragments into a suitable expression plasmid with a selectable marker; transforming the plasmids containing the DNA fragments into the organism from which the gene of interest originated; and selecting clones from transformed cells that show a phenotypic difference of the clone grown in the presence of the inducer relative to the phenotype in the absence of inducer.

    摘要翻译： 本发明提供了生产和选择药物敏化反义DNA片段的方法。 在一个实施方案中，该方法包括使用细菌生理学，生物化学，遗传学，基因组学和其他方法的知识来鉴定感兴趣的基因。 该方法包括使用基因组DNA作为模板对感兴趣的基因进行PCR扩增; 通过超声处理或其他方法破碎DNA; 选择不超过400个碱基对的DNA片段; 将DNA片段连接到具有选择性标记的合适的表达质粒中; 将含有DNA片段的质粒转化为目标基因来源的生物体; 并选择来自转化细胞的克隆，其在不存在诱导物的情况下显示在诱导物存在下生长的克隆相对于表型的表型差异。

    Method for identifying drug-sensitizing antisense DNA fragments and use thereof
    3.
    发明申请
    Method for identifying drug-sensitizing antisense DNA fragments and use thereof 有权
    用于鉴定药物敏化反义DNA片段的方法及其用途

    公开(公告)号：US20070218481A1

    公开(公告)日：2007-09-20

    申请号：US11636394

    申请日：2006-12-08

    申请人： Robert Haselbeck ,  Mark Hilgers ,  Karen Shaw ,  Vickie Brown-Driver ,  Kedar Gc ,  John M. Finn ,  Mark Stidham

    发明人： Robert Haselbeck ,  Mark Hilgers ,  Karen Shaw ,  Vickie Brown-Driver ,  Kedar Gc ,  John M. Finn ,  Mark Stidham

    IPC分类号： C12Q1/68 ,  C07H21/02 ,  C12N15/74 ,  C12N1/21

    CPC分类号： C12N15/111 ,  C12N15/1137 ,  C12N2310/11 ,  C12N2320/11 ,  C12N2330/30 ,  C12Y101/01158 ,  C12Y105/01003 ,  C12Y205/01031 ,  C12Y601/0101

    摘要： The invention provides a method for generating and selecting drug-sensitizing antisense DNA fragments. In one embodiment, the method includes identifying a gene of interest using knowledge of bacterial physiology, biochemistry, genetics, genomics, and other means. The method includes PCR amplification of a gene of interest using genomic DNA as a template; fragmentation of the DNA by sonication or other means; selecting DNA fragments no longer than 400 base pairs; ligating the DNA fragments into a suitable expression plasmid with a selectable marker; transforming the plasmids containing the DNA fragments into the organism from which the gene of interest originated; and selecting clones from transformed cells that show a phenotypic difference of the clone grown in the presence of the inducer relative to the phenotype in the absence of inducer.

    摘要翻译： 本发明提供了生产和选择药物敏化反义DNA片段的方法。 在一个实施方案中，该方法包括使用细菌生理学，生物化学，遗传学，基因组学和其他方法的知识来鉴定感兴趣的基因。 该方法包括使用基因组DNA作为模板对感兴趣的基因进行PCR扩增; 通过超声处理或其他方法破碎DNA; 选择不超过400个碱基对的DNA片段; 将DNA片段连接到具有选择性标记的合适的表达质粒中; 将含有DNA片段的质粒转化为目标基因来源的生物体; 并选择来自转化细胞的克隆，其在不存在诱导物的情况下显示在诱导物存在下生长的克隆相对于表型的表型差异。

    Identification and application of antibiotic synergy
    4.
    发明授权
    Identification and application of antibiotic synergy 有权
    鉴定和应用抗生素协同作用

    公开(公告)号：US08298543B2

    公开(公告)日：2012-10-30

    申请号：US13102856

    申请日：2011-05-06

    申请人： Vickie Brown-Driver ,  Kedar GC ,  John M. Finn ,  Robert Haselbeck ,  Mark Hilgers ,  Karen Shaw ,  Mark Stidham

    发明人： Vickie Brown-Driver ,  Kedar GC ,  John M. Finn ,  Robert Haselbeck ,  Mark Hilgers ,  Karen Shaw ,  Mark Stidham

    IPC分类号： A61K31/00 ,  A61K31/43 ,  A61K39/00

    CPC分类号： A61K31/43 ,  A61K31/545 ,  A61K39/40 ,  A61K45/06 ,  Y02A50/469 ,  Y02A50/47 ,  Y02A50/482 ,  Y02A50/484 ,  A61K2300/00

    摘要： The present invention provides a pharmaceutical composition useful for treating bacterial infections in humans and animals which comprises administering to a human or animal in need thereof, an antibacterially effective combination of a β-lactam antibiotic and an inhibitor of any bacterial peptidoglycan biosynthesis enzyme, especially GlmU, GlmU, MurA, MurB, MurC, MurD, MurE, MurF, MurG, MraY, and UppS. Further provided is a method of discovering synergists for antibiotics including: a) expressing in a cell an antisense nucleic acid against a nucleic acid encoding a gene product so as to reduce the activity or amount of the gene product in the cell, thereby producing a cell sensitized to an antibiotic; b) characterizing the sensitization of the cell to the antibiotic and selecting pairs of antibiotics and genes that result in antibiotic efficacy at one-fifth or less the concentration required in the absence of the antisense gene; c) screening for chemical compounds that inhibit the gene product corresponding to the selected synergistic gene; and d) selecting or creating chemical analogs that inhibit the gene product corresponding to the selected synergistic gene such that the inhibition occurs in the bacteria.

    摘要翻译： 本发明提供了一种可用于治疗人和动物细菌感染的药物组合物，其包括对有需要的人或动物施用抗生素有效组合的β-内酰胺抗生素和任何细菌肽聚糖生物合成酶的抑制剂，特别是 GlmU，GlmU，MurA，MurB，MurC，MurD，MurE，MurF，MurG，MraY，和UppS。 进一步提供了发现抗生素增效剂的方法，包括：a）在细胞中表达反义核酸对编码基因产物的核酸，以降低细胞中基因产物的活性或数量，从而产生细胞 对抗生素敏感; b）表征细胞对抗生素的致敏性，并选择一组抗生素和基因，其抗生素效力为不存在反义基因所需浓度的五分之一或更低。 c）筛选抑制与所选协同基因相对应的基因产物的化合物; 和d）选择或产生抑制与所选择的协同基因相对应的基因产物的化学类似物，使得在细菌中发生抑制。

    IDENTIFICATION AND APPLICATION OF ANTIBIOTIC SYNERGY
    5.
    发明申请
    IDENTIFICATION AND APPLICATION OF ANTIBIOTIC SYNERGY 有权
    抗生素合成的鉴定与应用

    公开(公告)号：US20110294774A1

    公开(公告)日：2011-12-01

    申请号：US13102856

    申请日：2011-05-06

    申请人： Vickie Brown-Driver ,  Kedar GC ,  John M. Finn ,  Robert Haselbeck ,  Mark Hilgers ,  Karen Shaw ,  Mark Stidham

    发明人： Vickie Brown-Driver ,  Kedar GC ,  John M. Finn ,  Robert Haselbeck ,  Mark Hilgers ,  Karen Shaw ,  Mark Stidham

    IPC分类号： A61K31/545 ,  A61P31/04 ,  A61K31/43 ,  A61K31/431 ,  A61K31/546 ,  A61K31/5365

    CPC分类号： A61K31/43 ,  A61K31/545 ,  A61K39/40 ,  A61K45/06 ,  Y02A50/469 ,  Y02A50/47 ,  Y02A50/482 ,  Y02A50/484 ,  A61K2300/00

    摘要： The present invention provides a pharmaceutical composition useful for treating bacterial infections in humans and animals which comprises administering to a human or animal in need thereof, an antibacterially effective combination of a β-lactam antibiotic and an inhibitor of any bacterial peptidoglycan biosynthesis enzyme, especially GlmU, GlmU, MurA, MurB, MurC, MurD, MurE, MurF, MurG, MraY, and UppS. Further provided is a method of discovering synergists for antibiotics including: a) expressing in a cell an antisense nucleic acid against a nucleic acid encoding a gene product so as to reduce the activity or amount of the gene product in the cell, thereby producing a cell sensitized to an antibiotic; b) characterizing the sensitization of the cell to the antibiotic and selecting pairs of antibiotics and genes that result in antibiotic efficacy at one-fifth or less the concentration required in the absence of the antisense gene; c) screening for chemical compounds that inhibit the gene product corresponding to the selected synergistic gene; and d) selecting or creating chemical analogs that inhibit the gene product corresponding to the selected synergistic gene such that the inhibition occurs in the bacteria.

    摘要翻译： 本发明提供了一种可用于治疗人和动物细菌感染的药物组合物，其包括对有需要的人或动物施用抗生素有效组合的β-内酰胺抗生素和任何细菌肽聚糖生物合成酶的抑制剂，特别是 GlmU，GlmU，MurA，MurB，MurC，MurD，MurE，MurF，MurG，MraY和UppS。 进一步提供了发现抗生素增效剂的方法，包括：a）在细胞中表达反义核酸对抗编码基因产物的核酸，以降低细胞中基因产物的活性或数量，从而产生细胞 对抗生素敏感; b）表征细胞对抗生素的致敏性，并选择一组抗生素和基因，其抗生素效力为不存在反义基因所需浓度的五分之一或更低。 c）筛选抑制与所选协同基因相对应的基因产物的化合物; 和d）选择或产生抑制与所选择的协同基因相对应的基因产物的化学类似物，使得在细菌中发生抑制。

    Identification and application of antibiotic synergy
    6.
    发明授权
    Identification and application of antibiotic synergy 有权
    鉴定和应用抗生素协同作用

    公开(公告)号：US08021665B2

    公开(公告)日：2011-09-20

    申请号：US11636379

    申请日：2006-12-08

    申请人： Vickie Brown-Driver ,  Kedar GC ,  John M. Finn ,  Robert Haselbeck ,  Mark Hilgers ,  Karen Shaw ,  Mark Stidham

    发明人： Vickie Brown-Driver ,  Kedar GC ,  John M. Finn ,  Robert Haselbeck ,  Mark Hilgers ,  Karen Shaw ,  Mark Stidham

    IPC分类号： A61K31/00 ,  A61K31/43 ,  A61K39/00

    CPC分类号： A61K31/43 ,  A61K31/545 ,  A61K39/40 ,  A61K45/06 ,  Y02A50/469 ,  Y02A50/47 ,  Y02A50/482 ,  Y02A50/484 ,  A61K2300/00

    摘要： The present invention provides a pharmaceutical composition useful for treating bacterial infections in humans and animals which comprises administering to a human or animal in need thereof, an antibacterially effective combination of a β-lactam antibiotic and an inhibitor of any bacterial peptidoglycan biosynthesis enzyme, especially GlmU, GlmU, MurA, MurB, MurC, MurD, MurE, MurF, MurG, MraY, and UppS. Further provided is a method of discovering synergists for antibiotics including: a) expressing in a cell an antisense nucleic acid against a nucleic acid encoding a gene product so as to reduce the activity or amount of the gene product in the cell, thereby producing a cell sensitized to an antibiotic; b) characterizing the sensitization of the cell to the antibiotic and selecting pairs of antibiotics and genes that result in antibiotic efficacy at one-fifth or less the concentration required in the absence of the antisense gene; c) screening for chemical compounds that inhibit the gene product corresponding to the selected synergistic gene; and d) selecting or creating chemical analogs that inhibit the gene product corresponding to the selected synergistic gene such that the inhibition occurs in the bacteria.

    摘要翻译： 本发明提供了一种可用于治疗人和动物细菌感染的药物组合物，其包括对有需要的人或动物施用抗生素有效组合的β-内酰胺抗生素和任何细菌肽聚糖生物合成酶的抑制剂，特别是 GlmU，GlmU，MurA，MurB，MurC，MurD，MurE，MurF，MurG，MraY和UppS。 进一步提供了发现抗生素增效剂的方法，包括：a）在细胞中表达反义核酸对抗编码基因产物的核酸，以降低细胞中基因产物的活性或数量，从而产生细胞 对抗生素敏感; b）表征细胞对抗生素的致敏性，并选择一组抗生素和基因，其抗生素效力为不存在反义基因所需浓度的五分之一或更低。 c）筛选抑制与所选协同基因相对应的基因产物的化合物; 和d）选择或产生抑制与所选择的协同基因相对应的基因产物的化学类似物，使得在细菌中发生抑制。

    Identification and application of antibiotic synergy
    7.
    发明申请
    Identification and application of antibiotic synergy 有权
    鉴定和应用抗生素协同作用

    公开(公告)号：US20070178111A1

    公开(公告)日：2007-08-02

    申请号：US11636379

    申请日：2006-12-08

    申请人： Vickie Brown-Driver ,  Kedar GC ,  John Finn ,  Robert Haselbeck ,  Mark Hilgers ,  Karen Shaw ,  Mark Stidham

    发明人： Vickie Brown-Driver ,  Kedar GC ,  John Finn ,  Robert Haselbeck ,  Mark Hilgers ,  Karen Shaw ,  Mark Stidham

    IPC分类号： A61K48/00 ,  A61K39/40 ,  A61K31/43 ,  A61K31/545

    CPC分类号： A61K31/43 ,  A61K31/545 ,  A61K39/40 ,  A61K45/06 ,  Y02A50/469 ,  Y02A50/47 ,  Y02A50/482 ,  Y02A50/484 ,  A61K2300/00

    摘要： The present invention provides a pharmaceutical composition useful for treating bacterial infections in humans and animals which comprises administering to a human or animal in need thereof, an antibacterially effective combination of a β-lactam antibiotic and an inhibitor of any bacterial peptidoglycan biosynthesis enzyme, especially GlmU, GlmU, MurA, MurB, MurC, MurD, MurE, MurF, MurG, MraY, and UppS. Further provided is a method of discovering synergists for antibiotics including: a) expressing in a cell an antisense nucleic acid against a nucleic acid encoding a gene product so as to reduce the activity or amount of the gene product in the cell, thereby producing a cell sensitized to an antibiotic; b) characterizing the sensitization of the cell to the antibiotic and selecting pairs of antibiotics and genes that result in antibiotic efficacy at one-fifth or less the concentration required in the absence of the antisense gene; c) screening for chemical compounds that inhibit the gene product corresponding to the selected synergistic gene; and d) selecting or creating chemical analogs that inhibit the gene product corresponding to the selected synergistic gene such that the inhibition occurs in the bacteria.

    摘要翻译： 本发明提供了用于治疗人和动物细菌感染的药物组合物，其包括对有需要的人或动物施用β-内酰胺抗生素和任何细菌肽聚糖生物合成酶的抑制剂的抗菌有效组合，特别是GlmU ，GlmU，MurA，MurB，MurC，MurD，MurE，MurF，MurG，MraY和UppS。 进一步提供了发现抗生素增效剂的方法，包括：a）在细胞中表达反义核酸对抗编码基因产物的核酸，以降低细胞中基因产物的活性或数量，从而产生细胞 对抗生素敏感; b）表征细胞对抗生素的致敏性，并选择一组抗生素和基因，其抗生素效力为不存在反义基因所需浓度的五分之一或更低。 c）筛选抑制与所选协同基因相对应的基因产物的化合物; 和d）选择或产生抑制与所选择的协同基因相对应的基因产物的化学类似物，使得在细菌中发生抑制。