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    METHOD FOR IDENTIFYING ANTIBIOTIC TARGETS BY COMPLEMENTED SEQUENCING
    1.
    发明申请
    METHOD FOR IDENTIFYING ANTIBIOTIC TARGETS BY COMPLEMENTED SEQUENCING 有权
    通过完整序列鉴定抗生素目标的方法

    公开(公告)号：US20140135234A1

    公开(公告)日：2014-05-15

    申请号：US14071618

    申请日：2013-11-04

    Applicant: DISCUVA LIMITED

    Inventor： DAVID HUGH WILLIAMS ,  Arthur Keith Turner

    IPC: C12N15/10

    CPC classification number: C12N15/1082 ,  C12N15/102 ,  C40B30/06 ,  G06F19/18

    Abstract: Disclosed is a method for identifying an essential gene which serves as an antibiotic target in a bacterium, the method comprising the steps of: (a) generating an antibiotic resistant mutant of said bacterium by a method comprising the step of selecting for growth in the presence of said antibiotic to produce an antibiotic resistant mutant clone (AbR mutant); (b) transforming the AbR mutant with: (i) one or more essential genes of said bacterium; and (ii) a transposon which insertionally inactivates bacterial DNA, to produce a pool of transposon mutants which are merodiploid for said one or more essential genes; (c) growing bacteria from the merodiploid pool in the presence of different amounts of said antibiotic to produce two or more test cultures; and (d) comparing the distribution of transposon insertions between test cultures to identify a putative essential gene serving as a target of said antibiotic in said bacterium.

    Abstract translation: 公开了一种用于鉴定在细菌中作为抗生素靶标的必需基因的方法，所述方法包括以下步骤：（a）通过包括以下步骤的方法产生所述细菌的抗生素抗性突变体：在存在时选择生长的步骤 的抗生素以产生抗生素抗性突变体克隆（AbR突变体）; （b）用以下方法转化AbR突变体：（i）所述细菌的一种或多种必需基因; 和（ii）插入地使细菌DNA失活的转座子，以产生用于所述一种或多种必需基因的二聚体的转座子突变体库; （c）在存在不同量的所述抗生素的情况下从所述双二倍体池培养细菌以产生两种或更多种测试培养物; 和（d）比较测试培养物之间的转座子插入的分布，以鉴定作为所述细菌中所述抗生素靶标的推定的必需基因。

    Method for identifying antibiotic targets by complemented sequencing
    2.
    发明授权
    Method for identifying antibiotic targets by complemented sequencing 有权
    通过补体测序鉴定抗生素靶标的方法

    公开(公告)号：US09493770B2

    公开(公告)日：2016-11-15

    申请号：US14071618

    申请日：2013-11-04

    Applicant: DISCUVA LIMITED

    Inventor： David Hugh Williams ,  Arthur Keith Turner

    IPC: C40B30/06 ,  G06F19/18 ,  C12N15/10

    CPC classification number: C12N15/1082 ,  C12N15/102 ,  C40B30/06 ,  G06F19/18

    Abstract: Provided herein are methods, and related compositions for identifying a putative essential gene which serves as an antibiotic target in a bacterium, the method, in some embodiments, comprising (a) generating an antibiotic resistant mutant of the bacterium by a method comprising the step of selecting for growth in the presence of an antibiotic to produce an antibiotic resistant mutant clone (AbR mutant); (b) transforming the AbR mutant with one or more essential genes of the bacterium and a transposon which insertionally inactivates bacterial DNA to produce a pool of transposon mutants which are merodiploid for the one or more essential genes; (c) growing bacteria from the merodiploid pool in the presence of different amounts of the antibiotic to produce two or more test cultures; and (d) comparing the distribution of transposon insertions between the test cultures to identify a putative essential gene serving as a target of the antibiotic in the bacterium.

    Abstract translation: 本文提供了用于鉴定在细菌中用作抗生素靶标的推定必需基因的方法和相关组合物，在一些实施方案中，所述方法包括（a）通过包括以下步骤的方法产生所述细菌的抗生素抗性突变体： 在抗生素存在下选择生长以产生抗生素抗性突变体克隆（AbR突变体）; （b）用所述细菌的一种或多种必需基因转化所述AbR突变体，以及转座子，其插入地灭活细菌DNA以产生用于所述一种或多种必需基因的二聚体的转座子突变体库; （c）在不同量的抗生素存在下从单体二聚体培养细菌产生两种或更多种测试培养物; 和（d）比较测试培养物之间的转座子插入的分布，以鉴定用作细菌中抗生素靶标的推定的必需基因。

    Method for identifying antibiotic targets
    4.
    发明授权
    Method for identifying antibiotic targets 有权

    公开(公告)号：US09745572B2

    公开(公告)日：2017-08-29

    申请号：US14071615

    申请日：2013-11-04

    Applicant: DISCUVA LIMITED

    Inventor： David Hugh Williams ,  Arthur Keith Turner

    IPC: C12N15/10 ,  C12N1/21 ,  C40B30/06 ,  G06F19/18

    CPC classification number: C12N15/1082 ,  C12N15/102 ,  C40B30/06 ,  G06F19/18

    Abstract: Provided are methods for identifying differentially represented genes, the method comprising the steps of generating a pool of mutant bacteria by transposon mutagenesis with an activating transposon (TnA), wherein the TnA comprises a promoter such that transposon insertion into bacterial DNA increases the transcription of a gene at or near the insertion site; growing bacteria from the mutant pool in the presence of different amounts of said antibiotic to produce two or more test cultures; and comparing the distribution of TnA insertions between test cultures.

    METHOD FOR IDENTIFYING ANTIBIOTIC TARGETS
    7.
    发明申请
    METHOD FOR IDENTIFYING ANTIBIOTIC TARGETS 有权
    识别抗生素目标的方法

    公开(公告)号：US20140141979A1

    公开(公告)日：2014-05-22

    申请号：US14071615

    申请日：2013-11-04

    Applicant: DISCUVA LIMITED

    Inventor： DAVID HUGH WILLIAMS ,  Arthur Keith Turner

    IPC: C12N15/10

    CPC classification number: C12N15/1082 ,  C12N15/102 ,  C40B30/06 ,  G06F19/18

    Abstract: Disclosed is a method for identifying an essential gene which serves as an antibiotic target in a bacterium, the method comprising the steps of: (a) generating a pool of mutant bacteria by transposon mutagenesis with an activating transposon (TnA), wherein the TnA comprises a promoter such that transposon insertion into bacterial DNA increases the transcription of a gene at or near the insertion site; (b) growing bacteria from the mutant pool in the presence of different amounts of said antibiotic to produce two or more test cultures; and (c) comparing the distribution of TnA insertions between test cultures to identify a putative essential gene serving as a target of said antibiotic in said bacterium.

    Abstract translation: 公开了一种用于鉴定在细菌中作为抗生素靶标的必需基因的方法，所述方法包括以下步骤：（a）通过用活化转座子（TnA）通过转座子诱变产生突变细菌池，其中TnA包含 使转座子插入细菌DNA的启动子增加插入位点处或附近的基因的转录; （b）在存在不同量的所述抗生素的情况下从突变池中培养细菌以产生两种或更多种测试培养物; 和（c）比较测试培养物之间的TnA插入的分布，以鉴定用作所述细菌中所述抗生素靶标的推定的必需基因。

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