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    Method for increasing the antibiotic-producing ability of antibiotic-producing microorganisms
    1.
    发明授权
    Method for increasing the antibiotic-producing ability of antibiotic-producing microorganisms 失效
    增加产生抗生素的微生物的抗生素产生能力的方法

    公开(公告)号：US5672497A

    公开(公告)日：1997-09-30

    申请号：US575843

    申请日：1995-12-21

    申请人： Karen L. Cox ,  Scott E. Fishman ,  Charles L. Hershberger ,  Eugene T. Seno

    发明人： Karen L. Cox ,  Scott E. Fishman ,  Charles L. Hershberger ,  Eugene T. Seno

    IPC分类号： C12P19/62 ,  C12N15/00 ,  C07H21/04 ,  C12N15/09

    CPC分类号： C12P19/62

    摘要： A method for increasing the antibiotic-producing ability of an antibiotic-producing microbial host cell is disclosed. The method involves transforming an antibiotic-producing microorganism with a recombinant DNA cloning vector that codes for the expression of an antibiotic biosynthetic enzyme or other gene product. The gene preferably codes for a product that is rate-limiting in the antibiotic biosynthetic pathway in Streptomyces. Plasmids that are useful for increasing tylosin production in accordance with the present method include plasmids pHJL280, pHJL284, pHJL309, pHJL311, and pHJL315. The invention further comprises microorganisms transformed with plasmids pHJL280, pHJL284, pHJL309, pHJL311, and pHJL315 and also other microorganisms and vectors used in accordance with the present method.

    摘要翻译： 公开了提高产生抗生素的微生物宿主细胞的抗生素产生能力的方法。 该方法包括用编码抗生素生物合成酶或其他基因产物表达的重组DNA克隆载体转化产生抗生素的微生物。 该基因优选编码在链霉菌中的抗生素生物合成途径中限速的产物。 根据本方法可用于增加泰乐菌素生产的质粒包括质粒pHJL280，pHJL284，pHJL309，pHJL311和pHJL315。 本发明还包括用质粒pHJL280，pHJL284，pHJL309，pHJL311和pHJL315转化的微生物以及根据本方法使用的其它微生物和载体。

    Method for stabilizing and selecting recombinant DNA containing host cell
    4.
    发明授权
    Method for stabilizing and selecting recombinant DNA containing host cell 失效
    用于稳定和选择含有宿主细胞的重组DNA的方法

    公开(公告)号：US4650761A

    公开(公告)日：1987-03-17

    申请号：US550167

    申请日：1983-11-09

    申请人： Charles L. Hershberger ,  Paul R. Rosteck, Jr.

    发明人： Charles L. Hershberger ,  Paul R. Rosteck, Jr.

    IPC分类号： C07K14/575 ,  C07K14/62 ,  C12N15/63 ,  C12N15/70 ,  C12N15/00 ,  C07H21/04 ,  C12N1/00 ,  C12N1/20 ,  C12P19/34 ,  C12P21/00 ,  C12P21/02 ,  C12P21/04 ,  C12R1/19

    CPC分类号： C12N15/635 ,  C07K14/57581 ,  C07K14/62 ,  C12N15/70 ,  Y10S435/849 ,  Y10S930/18

    摘要： The present invention is an improvement in the method for stabilizing and selecting host cells containing recombinant DNA. The method involves transforming a host cell with a recombinant DNA cloning vector which contains the .about.2.5 kb BglII cI repressor gene-containing restriction fragment of bacteriophage .lambda. and then lysogenizing the transformed host cell with a lysogenic organism which contains a marker which is lethal or conditionally lethal in the host cell but which is repressed in the transformed host cell by the repressor gene contained in the recombinant DNA cloning vector. The vector additionally contains a gene which codes for the expression of human proinsulin. The invention further comprises related recombinant DNA cloning vectors, transformed host cells and lysogenized transformed host cells.

    摘要翻译： 本发明是用于稳定和选择含有重组DNA的宿主细胞的方法的改进。 该方法包括用含有含噬菌体λ的DIFFERENCE 2.5kb BglII cI阻遏物基因限制性片段的重组DNA克隆载体转化宿主细胞，然后用含有致死或有条件的标记物的溶原性生物溶解转化的宿主细胞 在宿主细胞中致死，但是通过重组DNA克隆载体中包含的阻遏物基因在转化的宿主细胞中被抑制。 该载体还含有编码人胰岛素原表达的基因。 本发明还包括相关的重组DNA克隆载体，转化的宿主细胞和溶解的转化的宿主细胞。

    Method of cloning modified streptomycetes DNA
    6.
    发明授权
    Method of cloning modified streptomycetes DNA 失效
    克隆修饰链霉菌DNA的方法

    公开(公告)号：US4710466A

    公开(公告)日：1987-12-01

    申请号：US654063

    申请日：1984-09-25

    申请人： Charles L. Hershberger ,  Jeffrey L. Larson

    发明人： Charles L. Hershberger ,  Jeffrey L. Larson

    IPC分类号： C12N15/09 ,  C12N1/20 ,  C12N15/63 ,  C12N15/76 ,  C12R1/19 ,  C12R1/465 ,  C12P19/34 ,  C12N1/00 ,  C12N15/00

    CPC分类号： C12N15/76 ,  C12N15/63 ,  Y10S435/886

    摘要： A method of cloning endogenously modified Streptomycetes DNA, which is normally rejected by restrictionless heterospecific hosts, is disclosed. The method uses bacteriophage lambda to construct a genomic library of modified Streptomycetes DNA; such lambda-containing Streptomycetes DNA is replicated to provide a source of non-modified Streptomycetes DNA. This non-modified DNA is subcloned into a selectable cloning vector and used to transform restrictionless hetero-specific hosts. The transformants can then be screened for clones containing genes of interest.

    摘要翻译： 公开了内源性修饰的链霉菌DNA的克隆方法，其通常被无限异种宿主排除。 该方法使用噬菌体λ构建修饰链霉菌DNA的基因组文库; 这样的含有λ的链霉菌DNA被复制以提供非修饰链霉菌DNA的来源。 将该未修饰的DNA亚克隆到可选择的克隆载体中，并用于转化无限制的异源特异性宿主。 然后可以转化转化体用于含有感兴趣基因的克隆。

    Ultrahigh copy number streptomycetes plasmids
    10.
    发明授权
    Ultrahigh copy number streptomycetes plasmids 失效
    超高拷贝数链霉菌质粒

    公开(公告)号：US4898828A

    公开(公告)日：1990-02-06

    申请号：US841920

    申请日：1986-03-20

    申请人： Charles L. Hershberger ,  Jeffrey L. Larson ,  Patricia A. Reynolds

    发明人： Charles L. Hershberger ,  Jeffrey L. Larson ,  Patricia A. Reynolds

    IPC分类号： C12N1/21 ,  C12N15/69 ,  C12N15/76

    CPC分类号： C12N15/76 ,  C12N15/69

    摘要： The present invention discloses an .about.1.4 kb Bc1I-Sau3A restriction fragment containing the minimal replicon of the Streptomyces coelicolor plasmid SCP2*. This minimal replicon was deduced from plasmid derivatives constructed by in vitro deletions and has been identified as the smallest, self-replicating segment of the SCP2* plasmid. The minimal replicon specifies an ultrahigh level of plasmid DNA (more than 1000 fold increase from the basic replicon) when vectors containing this sequence are transformed into Streptomyces lividans. This replicon can be used to construct smaller, more efficient recombinant DNA shuttle vectors to clone DNA into streptomycetes.

    摘要翻译： 本发明公开了含有天蓝链霉菌质粒SCP2 *的最小复制子的差异1.4kb Bc1I-Sau3A限制性片段。 通过体外缺失构建的质粒衍生物推导出该最小复制子，并且已被鉴定为SCP2 *质粒的最小的自我复制片段。 当含有该序列的载体转化成变色链霉菌（Streptomyces lividans）时，最小复制子指定超高水平的质粒DNA（从基本复制子增加1000倍以上）。 该复制子可用于构建更小，更有效的重组DNA穿梭载体将DNA克隆入链霉菌。