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公开(公告)号：US20110229938A1
公开(公告)日：2011-09-22
申请号：US12934630
申请日：2009-03-27
申请人： Eugenio Ferrari , Caroline M. Peres
发明人： Eugenio Ferrari , Caroline M. Peres
CPC分类号： C12N15/75
摘要： Certain aspects of this disclosure relate to a method of amplifying a genomic locus. In certain embodiments, the method may comprise: a) contacting a population of bacterial host cells with an inhibitor of an essential enzyme, where the bacterial host cells comprise a genomic locus of the structure: A1-P-M-A2, where A1 and A2 are direct repeats, P comprises a coding sequence for a polypeptide, and M comprises a coding sequence for the essential enzyme; and b) selecting for cells that are resistant to the inhibitor; wherein cells that are resistant to the inhibitor have multiple copies of the amplification unit.
摘要翻译： 本公开的某些方面涉及扩增基因组座位的方法。 在某些实施方案中，该方法可以包括：a）使细菌宿主细胞群与必需酶的抑制剂接触，其中细菌宿主细胞包含以下结构的基因组座位：A1-PM-A2，其中A1和A2为 直接重复，P包含多肽的编码序列，M包含必需酶的编码序列; 和b）选择抗抑制剂的细胞; 其中对抑制剂具有抗性的细胞具有多个拷贝的扩增单元。
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公开(公告)号：US09528114B2
公开(公告)日：2016-12-27
申请号：US12934630
申请日：2009-03-27
申请人： Eugenio Ferrari , Caroline M. Peres
发明人： Eugenio Ferrari , Caroline M. Peres
IPC分类号： C12N15/75
CPC分类号： C12N15/75
摘要： Certain aspects of this disclosure relate to a method of amplifying a genomic locus. In certain embodiments, the method may comprise: a) contacting a population of bacterial host cells with an inhibitor of an essential enzyme, where the bacterial host cells comprise a genomic locus of the structure: A1-P-M-A2, where A1 and A2 are direct repeats, P comprises a coding sequence for a polypeptide, and M comprises a coding sequence for the essential enzyme; and b) selecting for cells that are resistant to the inhibitor; wherein cells that are resistant to the inhibitor have multiple copies of the amplification unit.
摘要翻译： 本公开的某些方面涉及扩增基因组座位的方法。 在某些实施方案中，该方法可以包括：a）使细菌宿主细胞群与必需酶的抑制剂接触，其中细菌宿主细胞包含以下结构的基因组座位：A1-PM-A2，其中A1和A2是 直接重复，P包含多肽的编码序列，M包含必需酶的编码序列; 和b）选择抗抑制剂的细胞; 其中对抑制剂具有抗性的细胞具有多个拷贝的扩增单元。
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公开(公告)号：US20120183995A1
公开(公告)日：2012-07-19
申请号：US13356572
申请日：2012-01-23
IPC分类号： C12P21/00 , C12N1/21 , C12N9/50 , C12N9/42 , C12N9/14 , C12N9/24 , C12N9/20 , C12N9/90 , C12N9/10 , C12N9/12 , C12N15/75 , C12N9/26
摘要： The present invention provides cells that have been genetically manipulated to have an altered capacity to produce expressed proteins. In particular, the present invention relates to Gram-positive microorganisms, such as Bacillus species having enhanced expression of a protein of interest, wherein one or more chromosomal genes have been inactivated, and preferably wherein one or more chromosomal genes have been deleted from the Bacillus chromosome. In some further embodiments, one or more indigenous chromosomal regions have been deleted from a corresponding wild-type Bacillus host chromosome.
摘要翻译： 本发明提供已被遗传操作以具有改变产生表达蛋白质的能力的细胞。 特别地，本发明涉及革兰氏阳性微生物，例如具有增强的感兴趣蛋白表达的芽孢杆菌属物种，其中一个或多个染色体基因已被灭活，并且优选其中一个或多个染色体基因已经从芽孢杆菌中缺失 染色体。 在一些另外的实施方案中，一个或多个本征染色体区已经从相应的野生型芽孢杆菌宿主染色体中缺失。
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公开(公告)号：US08124399B2
公开(公告)日：2012-02-28
申请号：US10507720
申请日：2003-03-28
摘要： The present invention provides cells that have been genetically manipulated to have an altered capacity to produce expressed proteins. In particular, the present invention relates to Gram-positive microorganisms, such as Bacillus species having enhanced expression of a protein of interest, wherein one or more chromosomal genes have been inactivated, and preferably wherein one or more chromosomal genes have been deleted from the Bacillus chromosome. In some further embodiments, one or more indigenous chromosomal regions have been deleted from a corresponding wild-type Bacillus host chromosome.
摘要翻译： 本发明提供已被遗传操作以具有改变产生表达蛋白质的能力的细胞。 特别地，本发明涉及革兰氏阳性微生物，例如具有增强的感兴趣蛋白表达的芽孢杆菌属物种，其中一个或多个染色体基因已被灭活，并且优选其中一个或多个染色体基因已经从芽孢杆菌中缺失 染色体。 在一些另外的实施方案中，一个或多个本征染色体区已经从相应的野生型芽孢杆菌宿主染色体中缺失。
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公开(公告)号：US08293499B2
公开(公告)日：2012-10-23
申请号：US13377529
申请日：2012-02-20
CPC分类号： C12N9/16 , C07K14/32 , C12N9/1205
摘要： The present invention provides host cells that have been genetically manipulated to have an enhanced capacity to produce proteins of interest. In particular, the invention relates to modified Bacillus sp. Host cells that have at least one inactivated phr gene. The enhanced production of proteins of interest by the modified Bacillus sp. Host cells is further increased in modified Bacillus sp. Host cells that overexpress YmaH. Methods for producing proteins of interest in the modified host cells are also provided.
摘要翻译： 本发明提供已被遗传操作以具有增强的产生感兴趣蛋白质的能力的宿主细胞。 特别地，本发明涉及改良的芽孢杆菌属 具有至少一个失活的phr基因的宿主细胞。 通过改良的芽孢杆菌菌种增强生产的目的蛋白质 宿主细胞在改良的芽孢杆菌中进一步增加。 宿主细胞过度表达YmaH。 还提供了在修饰的宿主细胞中产生目标蛋白质的方法。
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公开(公告)号：US07544488B2
公开(公告)日：2009-06-09
申请号：US11725836
申请日：2007-03-19
IPC分类号： C12P21/06
CPC分类号： C07K14/32
摘要： The present invention relates to cells that have been genetically manipulated to have an altered capacity to produce expressed proteins. In particular, the present invention relates to Gram-positive microorganisms having exogenous nucleic acid sequences introduced therein and methods for producing proteins in such host cells, such as members of the genus Bacillus. More specifically, the present invention relates to the expression, production and secretion of a polypeptide of interest and to cells that have been genetically manipulated to have an altered capacity to produce expressed proteins. In particular, the present invention provides for the enhanced expression of a selected polypeptide by a microorganism.
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公开(公告)号：US20090075332A1
公开(公告)日：2009-03-19
申请号：US12047157
申请日：2008-03-12
申请人： Eugenio Ferrari , David A. Estell
发明人： Eugenio Ferrari , David A. Estell
CPC分类号： C12N9/54 , C12Y304/21062
摘要： This invention relates to modified polynucleotides encoding modified proteases, and methods for altering the production of proteases in microorganisms. In particular, the present invention relates to methods for altering the expression of proteases in microorganisms, such as Bacillus species. The invention discloses modified polynucleotides, vectors, modified polypeptides, and processes for enhancing the production of proteases.
摘要翻译： 本发明涉及编码修饰的蛋白酶的修饰多核苷酸，以及改变微生物中蛋白酶生成的方法。 特别地，本发明涉及用于改变微生物如芽孢杆菌属物种中蛋白酶表达的方法。 本发明公开了修饰的多核苷酸，载体，修饰的多肽以及用于增强蛋白酶生产的方法。
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公开(公告)号：US20060073559A1
公开(公告)日：2006-04-06
申请号：US10507720
申请日：2003-03-28
申请人： Eugenio Ferrari , Carole Harbison , M Rashid , Walter Weyler
发明人： Eugenio Ferrari , Carole Harbison , M Rashid , Walter Weyler
摘要： The present invention provides cells that have been genetically manipulated to have an altered capacity to produce expressed proteins. In particular, the present invention relates to Gram-positive microorganisms, such as Bacillus species having enhanced expression of a protein of interest, wherein one or more chromosomal genes have been inactivated, and preferably wherein one or more chromosomal genes have been deleted from the Bacillus chromosome. In some further embodiments, one or more indigenous chromosomal regions have been deleted from a corresponding wild-type Bacillus host chromosome.
摘要翻译： 本发明提供已被遗传操作以具有改变产生表达蛋白质的能力的细胞。 特别地，本发明涉及革兰氏阳性微生物，例如具有增强的感兴趣蛋白表达的芽孢杆菌属物种，其中一个或多个染色体基因已被灭活，并且优选其中一个或多个染色体基因已经从芽孢杆菌中缺失 染色体。 在一些另外的实施方案中，一个或多个本征染色体区已经从相应的野生型芽孢杆菌宿主染色体中缺失。
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公开(公告)号：US5264366A
公开(公告)日：1993-11-23
申请号：US797577
申请日：1991-11-25
申请人： Eugenio Ferrari , Dennis J. Henner , Mark L. Stahl
发明人： Eugenio Ferrari , Dennis J. Henner , Mark L. Stahl
CPC分类号： C12R1/07 , C11D3/386 , C12N15/102 , C12N15/75 , C12N9/54
摘要： There are described, normally sporulating mutant Bacillus strain(s) which produce no detectable proteolytic activity during any phase of its growth. The absence of detectable proteolytic activity is due to the deletion of one or more codons specifying the mature subtilisin protease and the mature neutral protease. Also described are methods for producing such normally sporulating, protease deficient Bacillus mutants.
摘要翻译： 描述了通常是孢子形成的突变芽孢杆菌菌株，其在其生长的任何阶段都不产生可检测的蛋白水解活性。 不存在可检测的蛋白水解活性是由于缺失一个或多个密码子，指定成熟的枯草杆菌蛋白酶和成熟的中性蛋白酶。 还描述了用于产生这种通常形成孢子的蛋白酶缺陷芽孢杆菌突变体的方法。
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公开(公告)号：US08383366B2
公开(公告)日：2013-02-26
申请号：US13356572
申请日：2012-01-23
摘要： The present invention provides cells that have been genetically manipulated to have an altered capacity to produce expressed proteins. In particular, the present invention relates to Gram-positive microorganisms, such as Bacillus species having enhanced expression of a protein of interest, wherein one or more chromosomal genes have been inactivated, and preferably wherein one or more chromosomal genes have been deleted from the Bacillus chromosome. In some further embodiments, one or more indigenous chromosomal regions have been deleted from a corresponding wild-type Bacillus host chromosome.
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