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    Benzoate-and anthranilate-inducible promoters
    4.
    发明授权
    Benzoate-and anthranilate-inducible promoters 失效
    苯甲酸盐和邻氨基苯甲酸盐诱导型启动子

    公开(公告)号：US07794972B2

    公开(公告)日：2010-09-14

    申请号：US11028156

    申请日：2005-01-03

    申请人： Diane M. Retallack ,  Venkiteswaran Subramanian

    发明人： Diane M. Retallack ,  Venkiteswaran Subramanian

    IPC分类号： C12P21/02 ,  C12N1/21 ,  C07H21/04

    CPC分类号： C07K14/21 ,  C12N15/78

    摘要： Novel benzoate- or anthranilate-inducible promoters, and novel tandem promoters, and variants and improved mutants thereof, useful for commercial prokaryotic fermentation systems, nucleic acid constructs containing the promoters, expression systems using them, methods for expressing proteins by use thereof, and proteins expressed thereby.

    摘要翻译： 可用于商业原核发酵系统的新型苯甲酸盐或邻氨基苯甲酸盐诱导型启动子和新型串联启动子及其变体和改良突变体，含有启动子的核酸构建体，使用它们的表达系统，使用它们表达蛋白质的方法和蛋白质 由此表达。

    Production of recombinant proteins utilizing non-antibiotic selection methods and the incorporation of non-natural amino acids therein
    6.
    发明授权
    Production of recombinant proteins utilizing non-antibiotic selection methods and the incorporation of non-natural amino acids therein 有权
    使用非抗生素选择方法生产重组蛋白，并在其中掺入非天然氨基酸

    公开(公告)号：US09243253B2

    公开(公告)日：2016-01-26

    申请号：US13319844

    申请日：2010-05-10

    申请人： Diane M. Retallack ,  Lawrence C. Chew ,  Charles H. Squires

    发明人： Diane M. Retallack ,  Lawrence C. Chew ,  Charles H. Squires

    IPC分类号： C12N15/00 ,  C12N15/65 ,  C12P21/02 ,  C12N15/78 ,  C12N15/63

    CPC分类号： C12N15/65 ,  C12N15/63 ,  C12N15/78 ,  C12P21/02

    摘要： Provided herein are methods and compositions for expression of a nucleic acid construct comprising nucleic acids encoding a) a recombinant polypeptide, and b) a prototrophy-restoring enzyme in a host cell that is auxotrophic for at least one metabolite. In various embodiments, the host cell is auxotrophic for a nitrogenous base compound or an amino acid. The invention involves introducing an analogue into the growth media for the host cell such that the analogue is incorporated into the recombinant polypeptide or a nucleic acid coding sequence thereof. In various embodiments, the compositions and methods disclosed herein result in improved recombinant protein expression compared to expression of recombinant protein in an antibiotic selection system, or compared to expression of the recombinant protein in an expression system that lacks a metabolite analogue.

    摘要翻译： 本文提供了用于表达包含编码a）重组多肽的核酸的核酸构建体的方法和组合物，以及b）对于至少一种代谢产物是营养缺陷型的宿主细胞中的原始营养恢复酶。 在各种实施方案中，宿主细胞对于含氮碱性化合物或氨基酸是营养缺陷型的。 本发明涉及将类似物引入宿主细胞的生长培养基中，使得类似物掺入重组多肽或其核酸编码序列中。 在各种实施方案中，与抗生素选择系统中的重组蛋白的表达相比，本文公开的组合物和方法导致改善的重组蛋白表达，或与缺乏代谢物类似物的表达系统中重组蛋白的表达进行比较。

    Method for Rapidly Screening Microbial Hosts to Identify Certain Strains with Improved Yield and/or Quality in the Expression of Heterologous Proteins
    7.
    发明申请
    Method for Rapidly Screening Microbial Hosts to Identify Certain Strains with Improved Yield and/or Quality in the Expression of Heterologous Proteins 有权
    快速筛选微生物宿主鉴定异源蛋白表达中产量和/或质量提高的某些菌株的方法

    公开(公告)号：US20100137162A1

    公开(公告)日：2010-06-03

    申请号：US12610207

    申请日：2009-10-30

    申请人： Diane M. Retallack ,  Charles H. Squires ,  Thomas M. Ramseier ,  Russell J. Coleman ,  Jane C. Schneider ,  Charies D. Hershberger

    发明人： Diane M. Retallack ,  Charles H. Squires ,  Thomas M. Ramseier ,  Russell J. Coleman ,  Jane C. Schneider ,  Charies D. Hershberger

    IPC分类号： C40B30/10 ,  C40B50/06 ,  C40B40/02

    CPC分类号： C12N15/1037 ,  C12N15/78

    摘要： The present invention provides an array for rapidly identifying a host cell population capable of producing heterologous protein with improved yield and/or quality. The array comprises one or more host cell populations that have been genetically modified to increase the expression of one or more target genes involved in protein production, decrease the expression of one or more target genes involved in protein degradation, or both. One or more of the strains in the array may express the heterologous protein of interest in a periplasm compartment, or may secrete the heterologous protein extracellularly through an outer cell wall. The strain arrays are useful for screening for improved expression of any protein of interest, including therapeutic proteins, hormones, a growth factors, extracellular receptors or ligands, proteases, kinases, blood proteins, chemokines, cytokines, antibodies and the like.

    摘要翻译： 本发明提供了一种用于快速鉴定能够产生具有改善的产量和/或质量的异源蛋白质的宿主细胞群的阵列。 该阵列包含已被遗传修饰的一个或多个宿主细胞群，以增加涉及蛋白质产生的一个或多个靶基因的表达，减少参与蛋白质降解的一个或多个靶基因的表达，或两者。 阵列中的一个或多个菌株可以在周质隔室中表达感兴趣的异源蛋白质，或者通过外细胞壁在细胞外分泌异源蛋白质。 菌株阵列可用于筛选任何目的蛋白质的表达，包括治疗性蛋白质，激素，生长因子，细胞外受体或配体，蛋白酶，激酶，血液蛋白，趋化因子，细胞因子，抗体等。

    Method for rapidly screening microbial hosts to identify certain strains with improved yield and/or quality in the expression of heterologous proteins
    8.
    发明授权
    Method for rapidly screening microbial hosts to identify certain strains with improved yield and/or quality in the expression of heterologous proteins 有权
    用于快速筛选微生物宿主以鉴定具有提高异源蛋白质表达的产量和/或质量的某些菌株的方法

    公开(公告)号：US09580719B2

    公开(公告)日：2017-02-28

    申请号：US12610207

    申请日：2009-10-30

    申请人： Diane M. Retallack ,  Charles H. Squires ,  Thomas M. Ramseier ,  Russell J. Coleman ,  Jane C. Schneider ,  Charles D. Hershberger

    发明人： Diane M. Retallack ,  Charles H. Squires ,  Thomas M. Ramseier ,  Russell J. Coleman ,  Jane C. Schneider ,  Charles D. Hershberger

    IPC分类号： C12N15/78

    CPC分类号： C12N15/1037 ,  C12N15/78

    摘要： The present invention provides an array for rapidly identifying a host cell population capable of producing a heterologous protein with improved yield and/or quality. The array comprises one or more host cell populations that have been genetically modified to increase the expression of one or more target genes involved in protein production, decrease the expression of one or more target genes involved in protein degradation, or both. One or more of the strains in the array may express the heterologous protein of interest in a periplasm compartment or may secrete the heterologous protein extracellularly through an outer cell wall. The strain arrays are useful for screening for improved expression of any protein of interest including therapeutic proteins, hormones, growth factors, extracellular receptors or ligands, proteases, kinases, blood proteins, chemokines, cytokines, antibodies and the like.

    摘要翻译： 本发明提供了一种用于快速鉴定能够产生具有改善的产量和/或质量的异源蛋白质的宿主细胞群的阵列。 该阵列包含已被遗传修饰的一个或多个宿主细胞群，以增加参与蛋白质产生的一个或多个靶基因的表达，减少参与蛋白质降解的一个或多个靶基因的表达，或两者。 阵列中的一个或多个菌株可以在周质隔室中表达感兴趣的异源蛋白质，或者通过外细胞壁在细胞外分泌异源蛋白质。 菌株阵列可用于筛选任何目的蛋白质的表达，包括治疗性蛋白质，激素，生长因子，细胞外受体或配体，蛋白酶，激酶，血液蛋白，趋化因子，细胞因子，抗体等。

    Production Of Recombinant Proteins Utilizing Non-Antibiotic Selection Methods And The Incorporation Of Non-Natural Amino Acids Therein
    9.
    发明申请
    Production Of Recombinant Proteins Utilizing Non-Antibiotic Selection Methods And The Incorporation Of Non-Natural Amino Acids Therein 有权
    生产使用非抗生素选择方法的重组蛋白质和其中非天然氨基酸的掺入

    公开(公告)号：US20120115187A1

    公开(公告)日：2012-05-10

    申请号：US13319844

    申请日：2010-05-10

    申请人： Diane M. Retallack

    发明人： Diane M. Retallack

    IPC分类号： C12P21/00

    CPC分类号： C12N15/65 ,  C12N15/63 ,  C12N15/78 ,  C12P21/02

    摘要： Provided herein are methods and compositions for expression of a nucleic acid construct comprising nucleic acids encoding a) a recombinant polypeptide, and b) a prototrophy-restoring enzyme in a host cell that is auxotrophic for at least one metabolite. In various embodiments, the host cell is auxotrophic for a nitrogenous base compound or an amino acid. The invention involves introducing an analogue into the growth media for the host cell such that the analogue is incorporated into the recombinant polypeptide or a nucleic acid coding sequence thereof. In various embodiments, the compositions and methods disclosed herein result in improved recombinant protein expression compared to expression of recombinant protein in an antibiotic selection system, or compared to expression of the recombinant protein in an expression system that lacks a metabolite analogue.

    摘要翻译： 本文提供了用于表达包含编码a）重组多肽的核酸的核酸构建体的方法和组合物，以及b）对于至少一种代谢产物是营养缺陷型的宿主细胞中的原始营养恢复酶。 在各种实施方案中，宿主细胞对于含氮碱性化合物或氨基酸是营养缺陷型的。 本发明涉及将类似物引入宿主细胞的生长培养基中，使得类似物掺入重组多肽或其核酸编码序列中。 在各种实施方案中，与抗生素选择系统中的重组蛋白的表达相比，本文公开的组合物和方法导致改善的重组蛋白表达，或与缺乏代谢物类似物的表达系统中重组蛋白的表达进行比较。

    Bacterial leader sequences for increased expression
    10.
    发明授权
    Bacterial leader sequences for increased expression 有权
    用于增加表达的细菌前导序列

    公开(公告)号：US07833752B2

    公开(公告)日：2010-11-16

    申请号：US12604061

    申请日：2009-10-22

    申请人： Russell J. Coleman ,  Diane M. Retallack ,  Stacey L. Lee ,  Charles D. Hershberger

    发明人： Russell J. Coleman ,  Diane M. Retallack ,  Stacey L. Lee ,  Charles D. Hershberger

    IPC分类号： C12P21/06 ,  C12N1/20 ,  C12N15/00 ,  C07K1/00

    CPC分类号： C12N15/625 ,  C07K2319/02 ,  C07K2319/036

    摘要： Compositions and methods for improving expression and/or secretion of a polypeptide of interest in a host cell are provided. Compositions including a coding sequence for a bacterial secretion signal peptide are provided. The compositions of the invention are useful for increasing accumulation of properly processed proteins in the periplasmic space of a host cell, or for increasing secretion of properly processed proteins. In particular, isolated secretion signal peptide-encoding nucleic acid molecules are provided. Additionally, amino acid sequences corresponding to the nucleic acid molecules are encompassed. The present invention provides for isolated nucleic acid molecules including nucleotide sequences encoding the amino acid sequences shown in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, and 24, and the nucleotide sequences set forth in SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, and 23, as well as variants and fragments thereof.

    摘要翻译： 提供用于改善宿主细胞中感兴趣的多肽的表达和/或分泌的组合物和方法。 提供了包括细菌分泌信号肽的编码序列的组合物。 本发明的组合物可用于增加适当加工的蛋白质在宿主细胞的周质空间中的积累，或用于增加适当加工的蛋白质的分泌。 特别地，提供了分离信号肽编码核酸分子。 另外，包含对应于核酸分子的氨基酸序列。 本发明提供了分离的核酸分子，包括编码SEQ ID NO：2,4,6,8,10,12,14,16,18,20,22和24所示的氨基酸序列的核苷酸序列， SEQ ID NO：1,3,5,7,9,11,13,15,17,19,21和23所示的核苷酸序列，以及其变体和片段。