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    Method for Stable Gene-Amplification in a Bacterial Host Cell
    2.
    发明申请
    Method for Stable Gene-Amplification in a Bacterial Host Cell 审中-公开
    细菌宿主细胞中稳定基因扩增的方法

    公开(公告)号：US20140178930A1

    公开(公告)日：2014-06-26

    申请号：US14180975

    申请日：2014-02-14

    Applicant: Novozymes A/S

    Inventor： Carsten Olsen ,  Michael Dolberg Rasmussen

    IPC: C12N15/75 ,  C12P21/00

    CPC classification number: C12N15/75 ,  C12N9/1007 ,  C12N9/2471 ,  C12P21/00 ,  C12Y201/01014 ,  C12Y302/01023

    Abstract: A bacterial host cell is disclosed including at least two copies of an amplification unit in its genome, the amplification unit including: i) at least one copy of a gene of interest, and ii) an expressible conditionally essential gene, wherein the conditionally essential gene is either promoterless or transcribed from a heterologous promoter having an activity substantially lower than the endogenous promoter of the conditionally essential gene, and wherein the conditionally essential gene if not functional would render the cell auxotrophic for at least one specific substance or unable to utilize one or more specific sole carbon source; methods for producing a protein using the cell of the invention, and methods for constructing the cell of the invention.

    Abstract translation: 公开了一种细菌宿主细胞，其包含在其基因组中的至少两个拷贝的扩增单元，所述扩增单元包括：i）目的基因的至少一个拷贝，和ii）可表达条件必需基因，其中所述条件必需基因 是无活性的或从具有基本上低于条件性必需基因的内源启动子的活性的异源启动子转录，并且其中条件必需基因如果不起作用将使细胞营养缺陷至少一种特定物质或不能利用一种或 更具体的唯一碳源; 使用本发明的细胞产生蛋白质的方法以及构建本发明的细胞的方法。

    Method for Stable Gene-Amplification in a Bacterial Host Cell
    4.
    发明申请
    Method for Stable Gene-Amplification in a Bacterial Host Cell 审中-公开
    细菌宿主细胞中稳定基因扩增的方法

    公开(公告)号：US20160362698A1

    公开(公告)日：2016-12-15

    申请号：US15252548

    申请日：2016-08-31

    Applicant: Novozymes A/S

    Inventor： Carsten Olsen ,  Michael Dolberg Rasmussen

    IPC: C12N15/75 ,  C12N9/38 ,  C12N9/10 ,  C12P21/00

    CPC classification number: C12N15/75 ,  C12N9/1007 ,  C12N9/2471 ,  C12P21/00 ,  C12Y201/01014 ,  C12Y302/01023

    Abstract: A bacterial host cell is disclosed including at least two copies of an amplification unit in its genome, the amplification unit including: i) at least one copy of a gene of interest, and ii) an expressible conditionally essential gene, wherein the conditionally essential gene is either promoterless or transcribed from a heterologous promoter having an activity substantially lower than the endogenous promoter of the conditionally essential gene, and wherein the conditionally essential gene if not functional would render the cell auxotrophic for at least one specific substance or unable to utilize one or more specific sole carbon source; methods for producing a protein using the cell of the invention, and methods for constructing the cell of the invention.

    Abstract translation: 公开了一种细菌宿主细胞，其包含在其基因组中的至少两个拷贝的扩增单元，所述扩增单元包括：i）目的基因的至少一个拷贝，和ii）可表达条件必需基因，其中所述条件必需基因 是无活性的或从具有基本上低于条件性必需基因的内源启动子的活性的异源启动子转录，并且其中条件必需基因如果不起作用将使细胞营养缺陷至少一种特定物质或不能利用一种或 更具体的唯一碳源; 使用本发明的细胞产生蛋白质的方法以及构建本发明的细胞的方法。

    Stable Genomic Integration of Multiple Polynucleotide Copies
    5.
    发明申请
    Stable Genomic Integration of Multiple Polynucleotide Copies 审中-公开
    多重多核苷酸复制的稳定基因组整合

    公开(公告)号：US20140273237A1

    公开(公告)日：2014-09-18

    申请号：US14221005

    申请日：2014-03-20

    Applicant: Novozymes A/S

    Inventor： Anne Breuner ,  Michael Dolberg Rasmussen

    IPC: C12N15/75

    CPC classification number: C12N15/75 ,  C12N15/1082 ,  C12N15/1086 ,  C12N15/90 ,  C12N15/902 ,  C12N2800/30 ,  C12N2830/50

    Abstract: Methods of constructing a cell comprising in its chromosome one or more copies of an open reading frame (ORF) or operon encoding at least one polypeptide of interest, each copy being under the transcriptional control of a heterologous promoter using a site specific recombinase and in vivo integration by recombination; means for carrying out the methods, resulting cells, and methods for producing a polypeptide of interest using the resulting cells.

    Abstract translation: 构建细胞的方法包括在其染色体中的一个或多个拷贝的开放阅读框（ORF）或编码至少一种目标多肽的操纵子，每个拷贝在异源启动子的转录控制下使用位点特异性重组酶和体内 通过重组整合; 用于使用所得细胞进行方法，所得细胞和用于产生感兴趣的多肽的方法的手段。

    TEMPERATURE-SENSITIVE RNA- Guided Endonuclease
    6.
    发明申请
    TEMPERATURE-SENSITIVE RNA- Guided Endonuclease 有权

    公开(公告)号：US20220275354A1

    公开(公告)日：2022-09-01

    申请号：US17611547

    申请日：2020-04-29

    Applicant: Novozymes A/S

    Inventor： Brian Koebmann ,  Michael Dolberg Rasmussen ,  Carsten Andersen

    IPC: C12N9/22 ,  C12N15/11 ,  C12N15/75 ,  C12N15/90

    Abstract: The present invention relates to temperature-sensitive variants of an RNA-guided endonuclease, polynucleotides encoding said variants, nucleic acid constructs and expression vectors comprising polynucleotides encoding said variants, host cells expressing said variant, methods of transient repression and expression of one or more DNA target sequence using said variants, and use of said variants, polynucleotides, nucleic acid constructs, expression vectors, host cells, and methods.

    TEMPERATURE-SENSITIVE CAS9 PROTEIN
    8.
    发明申请
    TEMPERATURE-SENSITIVE CAS9 PROTEIN 审中-公开

    公开(公告)号：US20200318086A1

    公开(公告)日：2020-10-08

    申请号：US16652920

    申请日：2018-11-07

    Applicant: Novozymes A/S

    Inventor： Carsten Anderson ,  Poul Erik Pedersen ,  Michael Dolberg Rasmussen ,  Carsten Hjort

    IPC: C12N9/22 ,  C12N15/63

    Abstract: The present invention relates to temperature-sensitive variants of a Class-II Cas9 protein, polynucleotides encoding said variants, nucleic acid constructs and expression vectors comprising polynucleotides encoding said variants, host cells expressing said variant, methods of transient repression and expression of one or more genome target sequence using said variants, and use of said variants, polynucleotides, nucleic acid constructs, expression vectors, host cells, and methods.

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