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公开(公告)号:US08137951B2
公开(公告)日:2012-03-20
申请号:US13084230
申请日:2011-04-11
申请人: Alan Sloma , Leslie Naggiar, legal representative , Regine Behr , William Widner , Maria Tang , David Sternberg , Linda Sternberg, legal representative , Stephen Brown
发明人: Alan Sloma , Regine Behr , William Widner , Maria Tang , David Sternberg , Stephen Brown
IPC分类号: C12N1/20 , C12N15/74 , C12N15/00 , C12P21/00 , C12P9/26 , C12P1/00 , C12N9/00 , C12N9/24 , C12N9/88 , C07H21/04
CPC分类号: C12P19/26 , C12N9/1051 , C12N15/52
摘要: The present invention relates to methods for producing a hyaluronic acid, comprising: (a) cultivating a Bacillus host cell under conditions suitable for production of the hyaluronic acid, wherein the Bacillus host cell comprises a nucleic acid construct comprising a hyaluronan synthase encoding sequence operably linked to a promoter sequence foreign to the hyaluronan synthase encoding sequence; and (b) recovering the hyaluronic acid from the cultivation medium. The present invention also relates to an isolated nucleic acid sequence encoding a hyaluronan synthase operon comprising a hyaluronan synthase gene and a UDP-glucose 6-dehydrogenase gene, and optionally one or more genes selected from the group consisting of a UDP-glucose pyrophosphorylase gene, UDP-N-acetylglucosamine pyrophosphorylase gene, and glucose-6-phosphate isomerase gene. The present invention also relates to isolated nucleic acid sequences encoding a UDP-glucose 6-dehydrogenase, UDP-glucose pyrophosphorylase, and UDP-N-acetylglucosamine pyrophosphorylase.
摘要翻译: 本发明涉及透明质酸的制备方法,其包括:(a)在适于产生透明质酸的条件下培养芽孢杆菌属宿主细胞,其中所述芽孢杆菌宿主细胞包含可操作地连接的透明质酸合酶编码序列的核酸构建体 涉及透明质酸合酶编码序列外源的启动子序列; 和(b)从培养基中回收透明质酸。 本发明还涉及编码包含透明质酸合酶基因和UDP-葡萄糖-6-脱氢酶基因的透明质酸合酶操纵子的分离的核酸序列,以及任选地一种或多种选自UDP-葡萄糖焦磷酸化酶基因, UDP-N-乙酰葡糖胺焦磷酸化酶基因和葡萄糖-6-磷酸异构酶基因。 本发明还涉及编码UDP-葡萄糖-6-脱氢酶,UDP-葡萄糖焦磷酸化酶和UDP-N-乙酰葡糖胺焦磷酸化酶的分离的核酸序列。
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公开(公告)号:US20110014662A1
公开(公告)日:2011-01-20
申请号:US12891548
申请日:2010-09-27
申请人: Alan Sloma , Leslie Naggiar , Regine Behr , William Widner , Maria Tang , David Sternberg , Linda Sternberg , Stephen Brown
发明人: Alan Sloma , Leslie Naggiar , Regine Behr , William Widner , Maria Tang , David Sternberg , Linda Sternberg , Stephen Brown
IPC分类号: C12P19/04
CPC分类号: C12P19/26 , C12N9/1051 , C12N15/52
摘要: The present invention relates to methods for producing a hyaluronic acid, comprising: (a) cultivating a Bacillus host cell under conditions suitable for production of the hyaluronic acid, wherein the Bacillus host cell comprises a nucleic acid construct comprising a hyaluronan synthase encoding sequence operably linked to a promoter sequence foreign to the hyaluronan synthase encoding sequence; and (b) recovering the hyaluronic acid from the cultivation medium. The present invention also relates to an isolated nucleic acid sequence encoding a hyaluronan synthase operon comprising a hyaluronan synthase gene and a UDP-glucose 6-dehydrogenase gene, and optionally one or more genes selected from the group consisting of a UDP-glucose pyrophosphorylase gene, UDP-N-acetylglucosamine pyrophosphorylase gene, and glucose-6-phosphate isomerase gene. The present invention also relates to isolated nucleic acid sequences encoding a UDP-glucose 6-dehydrogenase, UDP-glucose pyrophosphorylase, and UDP-N-acetylglucosamine pyrophosphorylase
摘要翻译: 本发明涉及透明质酸的制备方法,其包括:(a)在适于产生透明质酸的条件下培养芽孢杆菌属宿主细胞,其中所述芽孢杆菌宿主细胞包含可操作地连接的透明质酸合酶编码序列的核酸构建体 涉及透明质酸合酶编码序列外源的启动子序列; 和(b)从培养基中回收透明质酸。 本发明还涉及编码包含透明质酸合酶基因和UDP-葡萄糖-6-脱氢酶基因的透明质酸合酶操纵子的分离的核酸序列,以及任选的一种或多种选自UDP-葡萄糖焦磷酸化酶基因, UDP-N-乙酰葡糖胺焦磷酸化酶基因和葡萄糖-6-磷酸异构酶基因。 本发明还涉及编码UDP-葡萄糖-6-脱氢酶,UDP-葡萄糖焦磷酸化酶和UDP-N-乙酰葡糖胺焦磷酸化酶的分离的核酸序列
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公开(公告)号:US07811806B2
公开(公告)日:2010-10-12
申请号:US10326185
申请日:2002-12-20
申请人: Alan Sloma , Regine Behr , William Widner , Maria Tang , David Sternberg , Stephen Brown
发明人: Alan Sloma , Regine Behr , William Widner , Maria Tang , David Sternberg , Stephen Brown
CPC分类号: C12P19/26 , C12N9/1051 , C12N15/52
摘要: The present invention relates to methods for producing a hyaluronic acid, comprising: (a) cultivating a Bacillus host cell under conditions suitable for production of the hyaluronic acid, wherein the Bacillus host cell comprises a nucleic acid construct comprising a hyaluronan synthase encoding sequence operably linked to a promoter sequence foreign to the hyaluronan synthase encoding sequence; and (b) recovering the hyaluronic acid from the cultivation medium. The present invention also relates to an isolated nucleic acid sequence encoding a hyaluronan synthase operon comprising a hyaluronan synthase gene and a UDP-glucose 6-dehydrogenase gene, and optionally one or more genes selected from the group consisting of a UDP-glucose pyrophosphorylase gene, UDP-N-acetylglucosamine pyrophosphorylase gene, and glucose-6-phosphate isomerase gene. The present invention also relates to isolated nucleic acid sequences encoding a UDP-glucose 6-dehydrogenase, UDP-glucose pyrophosphorylase, and UDP-N-acetylglucosamine pyrophosphorylase.
摘要翻译: 本发明涉及透明质酸的制备方法,其包括:(a)在适于产生透明质酸的条件下培养芽孢杆菌属宿主细胞,其中所述芽孢杆菌宿主细胞包含可操作地连接的透明质酸合酶编码序列的核酸构建体 涉及透明质酸合酶编码序列外源的启动子序列; 和(b)从培养基中回收透明质酸。 本发明还涉及编码包含透明质酸合酶基因和UDP-葡萄糖-6-脱氢酶基因的透明质酸合酶操纵子的分离的核酸序列,以及任选地一种或多种选自UDP-葡萄糖焦磷酸化酶基因, UDP-N-乙酰葡糖胺焦磷酸化酶基因和葡萄糖-6-磷酸异构酶基因。 本发明还涉及编码UDP-葡萄糖-6-脱氢酶,UDP-葡萄糖焦磷酸化酶和UDP-N-乙酰葡糖胺焦磷酸化酶的分离的核酸序列。
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公开(公告)号:US5955310A
公开(公告)日:1999-09-21
申请号:US31442
申请日:1998-02-26
申请人: William Widner , Alan Sloma , Michael D. Thomas
发明人: William Widner , Alan Sloma , Michael D. Thomas
IPC分类号: C12N15/09 , C07K14/325 , C12N1/21 , C12N9/54 , C12N15/32 , C12N15/75 , C12R1/07 , C12P21/06 , C12N1/20 , C12N9/00 , C12N9/88
CPC分类号: C12N15/75
摘要: The present invention relates to methods for producing a polypeptide, comprising: (a) cultivating a Bacillus host cell in a medium conducive for the production of the polypeptide, wherein the Bacillus cell comprises a nucleic acid construct comprising (i) a tandem promoter in which each promoter sequence of the tandem promoter is operably linked to a single copy of a nucleic acid sequence encoding the polypeptide and (ii) an mRNA processing/stabilizing sequence located downstream of the tandem promoter and upstream of the nucleic acid sequence encoding the polypeptide; and (b) isolating the polypeptide from the cultivation medium. The present invention also relates to Bacillus cells for producing a polypeptide comprising a nucleic acid construct which comprises (i) a tandem promoter in which each promoter sequence of the tandem promoter is operably linked to a single copy of a nucleic acid sequence encoding the polypeptide and (ii) an mRNA processing/stabilizing sequence located downstream of the tandem promoter and upstream of the nucleic acid sequence encoding the polypeptide.
摘要翻译: 本发明涉及产生多肽的方法,其包括:(a)在有助于产生多肽的培养基中培养芽孢杆菌宿主细胞,其中所述芽孢杆菌细胞包含核酸构建体,其包含(i)串联启动子,其中 串联启动子的每个启动子序列可操作地连接到编码多肽的核酸序列的单拷贝,和(ii)位于串联启动子下游的mRNA加工/稳定序列和编码多肽的核酸序列的上游; 和(b)从培养基中分离多肽。 本发明还涉及用于产生包含核酸构建体的多肽的芽孢杆菌细胞,其包含(i)串联启动子,其中串联启动子的每个启动子序列可操作地连接到编码多肽的核酸序列的单拷贝,以及 (ii)位于串联启动子下游并编码该多肽的核酸序列上游的mRNA加工/稳定序列。
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公开(公告)号:US20110189737A1
公开(公告)日:2011-08-04
申请号:US13084230
申请日:2011-04-11
申请人: Alan Sloma , Regine Behr , William Widner , Maria Tang , David Sternberg , Stephen Brown , Leslie Naggiar , Linda Sternberg
发明人: Alan Sloma , Regine Behr , William Widner , Maria Tang , David Sternberg , Stephen Brown , Leslie Naggiar , Linda Sternberg
IPC分类号: C12P19/04
CPC分类号: C12P19/26 , C12N9/1051 , C12N15/52
摘要: The present invention relates to methods for producing a hyaluronic acid, comprising: (a) cultivating a Bacillus host cell under conditions suitable for production of the hyaluronic acid, wherein the Bacillus host cell comprises a nucleic acid construct comprising a hyaluronan synthase encoding sequence operably linked to a promoter sequence foreign to the hyaluronan synthase encoding sequence; and (b) recovering the hyaluronic acid from the cultivation medium. The present invention also relates to an isolated nucleic acid sequence encoding a hyaluronan synthase operon comprising a hyaluronan synthase gene and a UDP-glucose 6-dehydrogenase gene, and optionally one or more genes selected from the group consisting of a UDP-glucose pyrophosphorylase gene, UDP-N-acetylglucosamine pyrophosphorylase gene, and glucose-6-phosphate isomerase gene. The present invention also relates to isolated nucleic acid sequences encoding a UDP-glucose 6-dehydrogenase, UDP-glucose pyrophosphorylase, and UDP-N-acetylglucosamine pyrophosphorylase
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公开(公告)号:US06255076B1
公开(公告)日:2001-07-03
申请号:US09258377
申请日:1999-02-26
申请人: William Widner , Alan Sloma , Michael D. Thomas
发明人: William Widner , Alan Sloma , Michael D. Thomas
IPC分类号: C12P2106
CPC分类号: C12N15/75
摘要: The present invention relates to methods for producing a polypeptide, comprising: (a) cultivating a Bacillus host cell in a medium conducive for the production of the polypeptide, wherein the Bacillus cell comprises a nucleic acid construct comprising (i) a tandem promoter in which each promoter sequence of the tandem promoter is operably linked to a single copy of a nucleic acid sequence encoding the polypeptide and alternatively also (ii) an mRNA processing/stabilizing sequence located downstream of the tandem promoter and upstream of the nucleic acid sequence encoding the polypeptide; and (b) isolating the polypeptide from the cultivation medium. The present invention also relates to methods for producing a polypeptide, comprising: (a) cultivating a Bacillus host cell in a medium conducive for the production of the polypeptide, wherein the Bacillus cell comprises a nucleic acid construct comprising (i) a “consensus” promoter having the sequence TTGACA for the “−35” region and TATAAT for the “−10” region operably linked to a single copy of a nucleic acid sequence encoding the polypeptide and (ii) an mRNA processing/stabilizing sequence located downstream of the “consensus” promoter and upstream of the nucleic acid sequence encoding the polypeptide; and (b) isolating the polypeptide from the cultivation medium.
摘要翻译: 本发明涉及产生多肽的方法,其包括:(a)在有助于产生多肽的培养基中培养芽孢杆菌宿主细胞,其中所述芽孢杆菌细胞包含核酸构建体,其包含(i)串联启动子,其中 串联启动子的每个启动子序列可操作地连接到编码多肽的核酸序列的单拷贝,或者还有(ii)位于串联启动子下游的mRNA加工/稳定序列和编码多肽的核酸序列的上游 ; 和(b)从培养基中分离多肽。 本发明还涉及产生多肽的方法,其包括:(a)在有助于产生多肽的培养基中培养芽孢杆菌宿主细胞,其中所述芽孢杆菌细胞包含核酸构建体,其包含(i)“共有” 具有“-35”区域的TTGACA序列的启动子和“-10”区域的TATAAT可操作地连接到编码多肽的核酸序列的单拷贝,和(ii)位于“ 共有“启动子和编码多肽的核酸序列的上游; 和(b)从培养基中分离多肽。
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公开(公告)号:US3301450A
公开(公告)日:1967-01-31
申请号:US49210365
申请日:1965-10-01
申请人: WILLIAM WIDNER
发明人: WILLIAM WIDNER
IPC分类号: B62D43/02
CPC分类号: B62D43/02
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公开(公告)号:US08574886B2
公开(公告)日:2013-11-05
申请号:US13401663
申请日:2012-02-21
申请人: Regine Behr , William Widner , Maria Tang , Stephen Brown , Leslie Naggiar , Linda Sternberg
发明人: Alan Sloma , Regine Behr , William Widner , Maria Tang , David Sternberg , Stephen Brown
IPC分类号: C12N1/20 , C12N15/74 , C12N15/00 , C12P19/04 , C12P19/26 , C12P19/28 , C12P1/00 , C12P21/06 , C07H21/04
CPC分类号: C12P19/26 , C12N9/1051 , C12N15/52
摘要: The present invention relates to methods for producing a hyaluronic acid, comprising: (a) cultivating a Bacillus host cell under conditions suitable for production of the hyaluronic acid, wherein the Bacillus host cell comprises a nucleic acid construct comprising a hyaluronan synthase encoding sequence operably linked to a promoter sequence foreign to the hyaluronan synthase encoding sequence; and (b) recovering the hyaluronic acid from the cultivation medium. The present invention also relates to an isolated nucleic acid sequence encoding a hyaluronan synthase operon comprising a hyaluronan synthase gene and a UDP-glucose 6-dehydrogenase gene, and optionally one or more genes selected from the group consisting of a UDP-glucose pyrophosphorylase gene, UDP-N-acetylglucosamine pyrophosphorylase gene, and glucose-6-phosphate isomerase gene. The present invention also relates to isolated nucleic acid sequences encoding a UDP-glucose 6-dehydrogenase, UDP-glucose pyrophosphorylase, and UDP-N-acetylglucosamine pyrophosphorylase.
摘要翻译: 本发明涉及透明质酸的制备方法,其包括:(a)在适于产生透明质酸的条件下培养芽孢杆菌属宿主细胞,其中所述芽孢杆菌宿主细胞包含可操作地连接的透明质酸合酶编码序列的核酸构建体 涉及透明质酸合酶编码序列外源的启动子序列; 和(b)从培养基中回收透明质酸。 本发明还涉及编码包含透明质酸合酶基因和UDP-葡萄糖-6-脱氢酶基因的透明质酸合酶操纵子的分离的核酸序列,以及任选地一种或多种选自UDP-葡萄糖焦磷酸化酶基因, UDP-N-乙酰葡糖胺焦磷酸化酶基因和葡萄糖-6-磷酸异构酶基因。 本发明还涉及编码UDP-葡萄糖-6-脱氢酶,UDP-葡萄糖焦磷酸化酶和UDP-N-乙酰葡糖胺焦磷酸化酶的分离的核酸序列。
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公开(公告)号:US07179634B2
公开(公告)日:2007-02-20
申请号:US09834271
申请日:2001-04-12
申请人: William Widner , Alan Sloma , Michael D. Thomas
发明人: William Widner , Alan Sloma , Michael D. Thomas
CPC分类号: C12N15/75
摘要: The present invention relates to methods for producing a polypeptide, comprising: (a) cultivating a Bacillus host cell in a medium conducive for the production of the polypeptide, wherein the Bacillus cell comprises a nucleic acid construct comprising (i) a tandem promoter in which each promoter sequence of the tandem promoter is operably linked to a single copy of a nucleic acid sequence encoding the polypeptide and alternatively also (ii) an mRNA processing/stabilizing sequence located downstream of the tandem promoter and upstream of the nucleic acid sequence encoding the polypeptide; and (b) isolating the polypeptide from the cultivation medium. The present invention also relates to methods for producing a polypeptide, comprising: (a) cultivating a Bacillus host cell in a medium conducive for the production of the polypeptide, wherein the Bacillus cell comprises a nucleic acid construct comprising (i) a “consensus” promoter having the sequence TTGACA for the “−35” region and TATAAT for the “−10” region operably linked to a single copy of a nucleic acid sequence encoding the polypeptide and (ii) an mRNA processing/stabilizing sequence located downstream of the “consensus” promoter and upstream of the nucleic acid sequence encoding the polypeptide; and (b) isolating the polypeptide from the cultivation medium.
摘要翻译: 本发明涉及产生多肽的方法,其包括:(a)在有助于产生多肽的培养基中培养芽孢杆菌宿主细胞,其中所述芽孢杆菌细胞包含核酸构建体,其包含(i)串联启动子,其中 串联启动子的每个启动子序列可操作地连接到编码多肽的核酸序列的单拷贝,或者还有(ii)位于串联启动子下游的mRNA加工/稳定序列和编码多肽的核酸序列的上游 ; 和(b)从培养基中分离多肽。 本发明还涉及产生多肽的方法,其包括:(a)在有助于产生多肽的培养基中培养芽孢杆菌宿主细胞,其中所述芽孢杆菌细胞包含核酸构建体,其包含(i)“共有” 具有“-35”区域的TTGACA序列的启动子和“-10”区域的TATAAT可操作地连接到编码多肽的核酸序列的单拷贝,和(ii)位于“ 共有“启动子和编码多肽的核酸序列的上游; 和(b)从培养基中分离多肽。
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公开(公告)号:US20120301955A1
公开(公告)日:2012-11-29
申请号:US13584410
申请日:2012-08-13
申请人: Michael Thomas , Gloria Erichsen , William Widner
发明人: Michael Thomas , Gloria Erichsen , William Widner
IPC分类号: C12N15/113 , C12N15/63
摘要: The present invention relates to methods of producing a polypeptide having biological activity in a bacterial cell, comprising: (a) cultivating a bacterial host cell in a medium conducive for production of the polypeptide, wherein the bacterial host cell comprises a nucleic acid construct comprising a promoter region operably linked to a polynucleotide sequence encoding the polypeptide and a modified mRNA processing/stabilizing sequence located downstream of the promoter region and upstream of the ribosome binding site of the polynucleotide sequence encoding the polypeptide, wherein the modified mRNA processing/stabilizing sequence promotes higher expression of the polynucleotide sequence compared to an unmodified mRNA processing/stabilizing sequence; and (b) isolating the polypeptide having biological activity from the cultivation medium. The present invention also relates to such modified mRNA processing/stabilizing sequences, nucleic acid constructs, and bacterial host cells and to methods of obtaining such bacterial host cells.
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