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    Broad host range vectors for shotgun and expression library cloning in Gram negative bacteria
    3.
    发明授权
    Broad host range vectors for shotgun and expression library cloning in Gram negative bacteria 有权
    用于霰弹枪和表达文库克隆的革兰氏阴性细菌的广泛宿主范围载体

    公开(公告)号：US07846688B2

    公开(公告)日：2010-12-07

    申请号：US11505147

    申请日：2006-08-15

    申请人： Ryan Gill ,  Michael Lynch

    发明人： Ryan Gill ,  Michael Lynch

    IPC分类号： C12N15/00 ,  C12N15/10

    CPC分类号： C12N15/66

    摘要： The present invention concerns methods and compositions for the construction of a series of stable vectors for genomic library construction useful in Gram negative species. In certain embodiments, the vectors contain the pBBR1 replicon, capable of to stable replication in a broad range of Gram negative species. In various embodiments, the plasmid vectors may also contain bidirectional, rho-independent transcriptional terminators flanking the multiple cloning site, which allows for greater insert stability, and thus, greater genomic representation. Each vector may vary in its selection marker region, mobilization function, and promoter used to express insert sequences. These vectors are of use in the screening of highly representational genomic libraries in a broad variety of Gram negative species.

    摘要翻译： 本发明涉及用于构建用于革兰氏阴性物种的基因组文库构建的一系列稳定载体的方法和组合物。 在某些实施方案中，载体含有能够在宽范围的革兰氏阴性物种中稳定复制的pBBR1复制子。 在各种实施方案中，质粒载体还可以包含在多克隆位点侧翼的双向，独立于转录终止子，其允许更大的插入稳定性，因此更大的基因组表示。 每个载体可以在其选择标记区，动员功能和用于表达插入序列的启动子上变化。 这些载体可用于筛选广泛种类的革兰氏阴性物种中高度代表性的基因组文库。

    Method of differential display of prokaryotic messenger RNA by RTPCR
    5.
    发明授权
    Method of differential display of prokaryotic messenger RNA by RTPCR 失效
    通过RTPCR差异显示原核信使RNA的方法

    公开(公告)号：US06759195B1

    公开(公告)日：2004-07-06

    申请号：US09534366

    申请日：2000-03-24

    申请人： William E. Bentley ,  Ryan Gill

    发明人： William E. Bentley ,  Ryan Gill

    IPC分类号： C12Q168

    CPC分类号： C12N15/1096

    摘要： The present invention provides a method of differential display of prokaryotic messages RNA by RTPCR, the method comprising the steps of: adding a first primer mixture to a first nucleic acid sample including a first mixture of mRNA to form a first primer/first nucleic acid sample mixture; adding the first primer mixture to a second nucleic acid sample including a first mixture of mRNA to form a first primer/second nucleic acid sample mixture; incubating the first primer/first nucleic acid sample mixture to produce a first population of cDNA; incubating the first primer/second nucleic acid sample mixture to produce a second population of cDNA; adding a second primer to the first population of cDNA to form a second primer/first population of cDNA mixture; adding the second primer mixture to the second population of cDNA to form a second primer/second population of cDNA mixture; amplifying the second primer/first population of cDNA mixture to produce a third population of cDNA; amplifying the second primer/second population of cDNA mixture to produce a fourth population of cDNA; identifying the presence or level of mRNA in the third population of cDNA; and identifying the presence or level of mRNA in the fourth population of cDNA.

    摘要翻译： 本发明提供了通过RTPCR差异显示原核信息RNA的方法，所述方法包括以下步骤：将第一引物混合物加入到包含mRNA的第一混合物的第一核酸样品中以形成第一引物/第一核酸样品 混合物; 将第一引物混合物加入到包含mRNA的第一混合物的第二核酸样品中以形成第一引物/第二核酸样品混合物; 孵育第一引物/第一核酸样品混合物以产生第一个cDNA群; 孵育第一引物/第二核酸样品混合物以产生第二群cDNA; 向第一群cDNA加入第二引物以形成cDNA混合物的第二引物/第一群; 将第二引物混合物加入到第二群cDNA中以形成cDNA混合物的第二引物/第二群; 扩增cDNA混合物的第二引物/第一群体以产生cDNA的第三群; 扩增cDNA混合物的第二引物/第二群体以产生第四个cDNA群; 鉴定第三组cDNA中mRNA的存在或水平; 并鉴定第四组cDNA中mRNA的存在或水平。

    Broad host range vectors for shotgun and expression library cloning in Gram negative bacteria
    7.
    发明申请
    Broad host range vectors for shotgun and expression library cloning in Gram negative bacteria 有权
    用于霰弹枪和表达文库克隆的革兰氏阴性细菌的广泛宿主范围载体

    公开(公告)号：US20070059768A1

    公开(公告)日：2007-03-15

    申请号：US11505147

    申请日：2006-08-15

    申请人： Ryan Gill ,  Michael Lynch

    发明人： Ryan Gill ,  Michael Lynch

    IPC分类号： C40B50/06 ,  C12N15/74

    CPC分类号： C12N15/66

    摘要： The present invention concerns methods and compositions for the construction of a series of stable vectors for genomic library construction useful in Gram negative species. In certain embodiments, the vectors contain the pBBR1 replicon, capable of to stable replication in a broad range of Gram negative species. In various embodiments, the plasmid vectors may also contain bidirectional, rho-independent transcriptional terminators flanking the multiple cloning site, which allows for greater insert stability, and thus, greater genomic representation. Each vector may vary in its selection marker region, mobilization function, and promoter used to express insert sequences. These vectors are of use in the screening of highly representational genomic libraries in a broad variety of Gram negative species.

    摘要翻译： 本发明涉及用于构建用于革兰氏阴性物种的基因组文库构建的一系列稳定载体的方法和组合物。 在某些实施方案中，载体含有能够在宽范围的革兰氏阴性物种中稳定复制的pBBR1复制子。 在各种实施方案中，质粒载体还可以包含在多克隆位点侧翼的双向，独立于转录终止子，其允许更大的插入稳定性，因此更大的基因组表示。 每个载体可以在其选择标记区，动员功能和用于表达插入序列的启动子上变化。 这些载体可用于筛选广泛种类的革兰氏阴性物种中高度代表性的基因组文库。

    Mixed-library parallel gene mapping quantitative micro-array technique for genome-wide identification of trait conferring genes
    9.
    发明申请
    Mixed-library parallel gene mapping quantitative micro-array technique for genome-wide identification of trait conferring genes 有权
    混合库平行基因定位定量微阵列技术，用于全基因组鉴定赋予基因的性状

    公开(公告)号：US20060084098A1

    公开(公告)日：2006-04-20

    申请号：US11231018

    申请日：2005-09-20

    申请人： Ryan Gill ,  Michael Lynch ,  Tanya Warnecke

    发明人： Ryan Gill ,  Michael Lynch ,  Tanya Warnecke

    IPC分类号： C40B30/06 ,  C40B40/08

    CPC分类号： C07K14/245 ,  C12N15/1079

    摘要： The present disclosure concerns methods and compositions relating to mixed-library parallel gene trait mapping. In particular embodiments, the methods concern quantitative microarray hybridization techniques for genome-wide identification of trait conferring genes. In other embodiments, the compositions concern genetic elements that confer or are associated with a trait. In an exemplary embodiment, the trait is enhanced growth rate. In another exemplary embodiment, genetic elements that confer enhanced bacterial growth rate comprise part or all of the sequences of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4 or SEQ ID NO:5. In other embodiments, the genetic elements that confer enhanced bacterial growth rate correspond to the YliF, adrA, yeaP, yddV or ydeH genes of E. coli.

    摘要翻译： 本公开涉及与混合型库并行基因性状图谱相关的方法和组合物。 在具体实施方案中，该方法涉及定性微阵列杂交技术，用于全基因组鉴定赋予性状的基因。 在其它实施方案中，组合物涉及赋予或与性状相关的遗传元件。 在一个示例性实施例中，性状是增强的增长率。 在另一个示例性实施方案中，赋予增强的细菌生长速率的遗传元件包含SEQ ID NO：1，SEQ ID NO：2，SEQ ID NO：3，SEQ ID NO：4或SEQ ID NO：5的部分或全部序列 。 在其他实施方案中，赋予增强的细菌生长速率的遗传元件对应于大肠杆菌的YliF，adrA，yeaP，yddV或ydeH基因。