公开(公告)号：US06673613B2

公开(公告)日：2004-01-06

申请号：US09911781

申请日：2001-07-24

申请人： David L. Craft ,  C. Ron Wilson ,  Dudley Eirich ,  Yeyan Zhang

发明人： David L. Craft ,  C. Ron Wilson ,  Dudley Eirich ,  Yeyan Zhang

IPC分类号： C12N1581

CPC分类号： C12N15/81

摘要： A nucleic acid sequence including a CYP promoter operably linked to nucleic acid encoding a heterologous protein is provided to increase transcription of the nucleic acid. Expression vectors and host cells containing the nucleic acid sequence are also provided. The methods and compositions described herein are especially useful in the production of polycarboxylic acids by yeast cells.

摘要翻译： 提供了包含与编码异源蛋白质的核酸可操作地连接的CYP启动子的核酸序列，以增加核酸的转录。 还提供了含有核酸序列的表达载体和宿主细胞。 本文所述的方法和组合物特别可用于通过酵母细胞生产多元羧酸。

公开(公告)号：US09102989B2

公开(公告)日：2015-08-11

申请号：US12244677

申请日：2008-10-02

申请人： David L. Craft

发明人： David L. Craft

IPC分类号： C12N1/19 ,  C12R1/74 ,  C12P7/44

CPC分类号： C12R1/74 ,  C12P7/44

摘要： Genetically modified strains of C. tropicalis, which will not revert to wild-type activity at the POX 4 and/or POX 5 locus, are disclosed. The strains are β-oxidation blocked and have been transformed through homologous recombination with a construct which deletes a portion of the POX 4 and/or POX 5 gene. The modified strains may be used to increase yields of dicarboxylic acids produced in host cells of the strains. Methods for blocking the β-oxidation pathway in a C. tropicalis host cell are also provided.

摘要翻译： 披露了热带假丝酵母的转基因菌株，其不会在POX 4和/或POX 5基因座处恢复野生型活性。 菌株被阻断并通过与删除一部分POX 4和/或POX 5基因的构建体进行同源重组而转化。 修饰的菌株可用于增加菌株宿主细胞中产生的二羧酸的产量。 还提供了用于阻断热带假丝酵母宿主细胞中的氧化途径的方法。

公开(公告)号：US07160708B2

公开(公告)日：2007-01-09

申请号：US10418819

申请日：2003-04-18

申请人： L. Dudley Eirich ,  David L. Craft

发明人： L. Dudley Eirich ,  David L. Craft

IPC分类号： C12N9/02 ,  C12N15/00 ,  C12N1/20 ,  C12P21/04 ,  C12Q1/00 ,  C12Q1/34 ,  C07H21/04 ,  C12Q1/26 ,  C12Q1/68

CPC分类号： C12N9/0006

摘要： The present invention provides fatty alcohol oxidase (FAO) proteins and nucleic acid molecules encoding the FAO proteins. Also provided are analogs, derivatives, and enzymatically active fragments of the FAO proteins. Vectors and host cells comprising the nucleic acid molecules encoding the FAO proteins, analogs, derivatives and enzymatically active fragments thereof are also provided. In addition, FAO signature peptides and isolated nucleic acid molecules encoding the signature peptides are provided by the present invention. Methods of producing or increasing production of a subject FAO protein, methods for increasing aldehyde production during the second step of the ω-oxidation pathway of fatty acids, methods for increasing production of a ketone from an alcohol during the second step of the ω-oxidation pathway of fatty acids, and methods for increasing production of a dicarboxylic acid are also provided.

摘要翻译： 本发明提供脂肪醇氧化酶（FAO）蛋白质和编码FAO蛋白质的核酸分子。 还提供了FAO蛋白质的类似物，衍生物和酶活性片段。 还提供了包含编码FAO蛋白质，类似物，衍生物和酶活性片段的核酸分子的载体和宿主细胞。 此外，本发明提供了编码签名肽的FAO签名肽和分离的核酸分子。 生产或增加主题的方法FAO蛋白质，在脂肪酸的ω-氧化途径的第二步中增加醛生产的方法，在ω-氧化的第二步中从醇中增加酮的生产方法 还提供了脂肪酸的途径，以及增加二羧酸生产的方法。

公开(公告)号：US07063972B2

公开(公告)日：2006-06-20

申请号：US10138838

申请日：2002-05-03

申请人： C. Ron Wilson ,  David L. Craft ,  L. Dudley Eirich ,  Mark Eshoo ,  Krishna M. Madduri ,  Cathy A. Cornett ,  Alfred A. Brenner ,  Maria Tang ,  John C. Loper ,  Martin Gleeson

发明人： C. Ron Wilson ,  David L. Craft ,  L. Dudley Eirich ,  Mark Eshoo ,  Krishna M. Madduri ,  Cathy A. Cornett ,  Alfred A. Brenner ,  Maria Tang ,  John C. Loper ,  Martin Gleeson

IPC分类号： C12N9/02 ,  C12N15/00 ,  C12N1/20 ,  C12P21/04 ,  C07H21/04

CPC分类号： C12N9/0042 ,  C07K14/40 ,  C12N9/0071 ,  C12N9/0077 ,  C12P7/44

摘要： Novel genes have been isolated which encode cytochrome P450 and NADPH reductase enzymes of the ω-hydroxylase complex of C. tropicalis 20336. Vectors including these genes, transfected host cells and transformed host cells are provided. Methods of producing of cytochrome P450 and NADPH reductase enzymes are also provided which involve transforming a host cell with a gene encoding these enzymes and culturing the cells. Methods of increasing the production of a dicarboxylic acid and methods of increasing production of the aforementioned enzymes are also provided which involve increasing in the host cell the number of genes encoding these enzymes. A method for discriminating members of a gene family by quantifying the expression of genes is also provided.

摘要翻译： 已经分离出编码热带假丝酵母20336的ω-羟化酶复合物的细胞色素P450和NADPH还原酶的新型基因。提供了包括这些基因，转染的宿主细胞和转化的宿主细胞的载体。 还提供了生产细胞色素P450和NADPH还原酶的方法，其涉及用编码这些酶的基因转化宿主细胞并培养细胞。 还提供了增加二羧酸生产的方法和增加上述酶的生产的方法，其涉及在宿主细胞中增加编码这些酶的基因的数量。 还提供了通过定量基因表达来区分基因家族成员的方法。

公开(公告)号：US07049112B2

公开(公告)日：2006-05-23

申请号：US10139296

申请日：2002-05-03

申请人： C. Ron Wilson ,  David L. Craft ,  L. Dudley Eirich ,  Mark Eshoo ,  Krishna M. Madduri ,  Cathy A. Cornett ,  Alfred A. Brenner ,  Maria Tang ,  John C. Loper ,  Martin Gleeson

发明人： C. Ron Wilson ,  David L. Craft ,  L. Dudley Eirich ,  Mark Eshoo ,  Krishna M. Madduri ,  Cathy A. Cornett ,  Alfred A. Brenner ,  Maria Tang ,  John C. Loper ,  Martin Gleeson

IPC分类号： C12N9/02 ,  C12N15/00 ,  C12N1/20 ,  C07H21/04 ,  C12P21/04

CPC分类号： C12N9/0042 ,  C07K14/40 ,  C12N9/0071 ,  C12N9/0077 ,  C12P7/44

摘要： Novel genes have been isolated which encode cytochrome P450 and NADPH reductase enzymes of the ω-hydroxylase complex of C. tropicalis 20336. Vectors including these genes, transfected host cells and transformed host cells are provided. Methods of producing of cytochrome P450 and NADPH reductase enzymes are also provided which involve transforming a host cell with a gene encoding these enzymes and culturing the cells. Methods of increasing the production of a dicarboxylic acid and methods of increasing production of the aforementioned enzymes are also provided which involve increasing in the host cell the number of genes encoding these enzymes. A method for discriminating members of a gene family by quantifying the expression of genes is also provided.

摘要翻译： 已经分离出编码热带假丝酵母20336的ω-羟化酶复合物的细胞色素P450和NADPH还原酶的新型基因。提供了包括这些基因，转染的宿主细胞和转化的宿主细胞的载体。 还提供了生产细胞色素P450和NADPH还原酶的方法，其涉及用编码这些酶的基因转化宿主细胞并培养细胞。 还提供了增加二羧酸生产的方法和增加上述酶的生产的方法，其涉及在宿主细胞中增加编码这些酶的基因的数量。 还提供了通过定量基因表达来区分基因家族成员的方法。

公开(公告)号：US07388084B2

公开(公告)日：2008-06-17

申请号：US10640962

申请日：2003-08-14

申请人： C. Ron Wilson ,  David L. Craft ,  Yeyan Zhang ,  Jeffrey B. Stavenhagen

发明人： C. Ron Wilson ,  David L. Craft ,  Yeyan Zhang ,  Jeffrey B. Stavenhagen

IPC分类号： C07H21/02 ,  C07H21/04 ,  C12N1/20 ,  C12N15/00

CPC分类号： C12N9/0042

摘要： The present invention provides modified promoters from Candida troplicalis CYP and POX4 genes. The modified promoters have various sequence motifs added, deleted, or altered in order to modulate expression of a coding sequence operably linked thereto. The sequence motifs comprise repressors of gene induction (URS sequences) and activators of gene induction (UAS sequences) as well as oleic acid response elements (ORE sequences). Yeast host cells comprising such modified promoters are also provided. Methods of altering expression of a protein of the beta or omega oxidation pathways using a subject modified promoter are also provided.

摘要翻译： 本发明提供了来自假丝酵母CYP和POX4基因的修饰的启动子。 修饰的启动子具有添加，缺失或改变的各种序列基序，以便调节与其可操作地连接的编码序列的表达。 序列基序包括基因诱导阻遏物（URS序列）和基因诱导活化剂（UAS序列）以及油酸反应元件（ORE序列）。 还提供了包含这种修饰的启动子的酵母宿主细胞。 还提供了使用受试者修饰的启动子改变β或ω-氧化途径的蛋白质表达的方法。

公开(公告)号：US06790640B2

公开(公告)日：2004-09-14

申请号：US10400902

申请日：2003-03-31

申请人： David L. Craft ,  C. Ron Wilson ,  Dudley Eirich ,  Yeyan Zhang

发明人： David L. Craft ,  C. Ron Wilson ,  Dudley Eirich ,  Yeyan Zhang

IPC分类号： C12P2102

CPC分类号： C12N15/81

摘要： A nucleic acid sequence including a CYP promoter operably linked to nucleic acid encoding a heterologous protein is provided to increase transcription of the nucleic acid. Expression vectors and host cells containing the nucleic acid sequence are also provided. The methods and compositions described herein are especially useful in the production of polycarboxylic acids by yeast cells.

摘要翻译： 提供了包含与编码异源蛋白质的核酸可操作地连接的CYP启动子的核酸序列，以增加核酸的转录。 还提供了含有核酸序列的表达载体和宿主细胞。 本文所述的方法和组合物特别可用于通过酵母细胞生产多元羧酸。

公开(公告)号：US06331420B1

公开(公告)日：2001-12-18

申请号：US09302620

申请日：1999-04-30

申请人： C. Ron Wilson ,  David L. Craft ,  L. Dudley Eirich ,  Mark Eshoo ,  Krishna M. Madduri ,  Cathy A. Cornett ,  Alfred A. Brenner ,  Maria Tang ,  John C. Loper ,  Martin Gleeson

发明人： C. Ron Wilson ,  David L. Craft ,  L. Dudley Eirich ,  Mark Eshoo ,  Krishna M. Madduri ,  Cathy A. Cornett ,  Alfred A. Brenner ,  Maria Tang ,  John C. Loper ,  Martin Gleeson

IPC分类号： C12P746

CPC分类号： C12N9/0042 ,  C07K14/40 ,  C12N9/0071 ,  C12N9/0077 ,  C12P7/44

摘要： Novel genes have been isolated which encode cytochrome P450 and NADPH reductase enzymes of the &ohgr;-hydroxylase complex of C. tropicalis 20336. Vectors including these genes, transfected host cells and transformed host cells are provided. Methods of producing of cytochrome P450 and NADPH reductase enzymes are also provided which involve transforming a host cell with a gene encoding these enzymes and culturing the cells. Methods of increasing the production of a dicarboxylic acid and methods of increasing production of the aforementioned enzymes are also provided which involve increasing in the host cell the number of genes encoding these enzymes. A method for discriminating members of a gene family by quantifying the expression of genes is also provided.

摘要翻译： 已经分离出编码热带假丝酵母20336的ω-羟化酶复合物的细胞色素P450和NADPH还原酶的新型基因。提供了包括这些基因，转染的宿主细胞和转化的宿主细胞的载体。 还提供了生产细胞色素P450和NADPH还原酶的方法，其涉及用编码这些酶的基因转化宿主细胞并培养细胞。 还提供了增加二羧酸生产的方法和增加上述酶的生产的方法，其涉及在宿主细胞中增加编码这些酶的基因的数量。 还提供了通过定量基因表达来区分基因家族成员的方法。

公开(公告)号：US20100167361A1

公开(公告)日：2010-07-01

申请号：US12244677

申请日：2008-10-02

申请人： David L. Craft

发明人： David L. Craft

IPC分类号： C12P7/44 ,  C12N1/15

CPC分类号： C12R1/74 ,  C12P7/44

摘要： Genetically modified strains of C. tropicalis, which will not revert to wild-type activity at the POX 4 and/or POX 5 locus, are disclosed. The strains are β-oxidation blocked and have been transformed through homologous recombination with a construct which deletes a portion of the POX 4 and/or POX 5 gene. The modified strains may be used to increase yields of dicarboxylic acids produced in host cells of the strains. Methods for blocking the β-oxidation pathway in a C. tropicalis host cell are also provided.

摘要翻译： 披露了热带假丝酵母的转基因菌株，其不会在POX 4和/或POX 5基因座处恢复野生型活性。 菌株被阻断并通过与删除一部分POX 4和/或POX 5基因的构建体进行同源重组而转化。 修饰的菌株可用于增加菌株宿主细胞中产生的二羧酸的产量。 还提供了用于阻断热带假丝酵母宿主细胞中的氧化途径的方法。

公开(公告)号：US07109009B2

公开(公告)日：2006-09-19

申请号：US10405660

申请日：2003-04-02

申请人： C. Ron Wilson ,  David L. Craft ,  L. Dudley Eirich ,  Mark Eshoo ,  Krishna M. Madduri ,  Cathy A. Cornett ,  Alfred A. Brenner ,  Maria Tang ,  John C. Loper ,  Martin Gleeson

发明人： C. Ron Wilson ,  David L. Craft ,  L. Dudley Eirich ,  Mark Eshoo ,  Krishna M. Madduri ,  Cathy A. Cornett ,  Alfred A. Brenner ,  Maria Tang ,  John C. Loper ,  Martin Gleeson

IPC分类号： C12N9/02 ,  C12N1/20 ,  C12N15/00 ,  C12Q1/00 ,  C12Q1/68 ,  C07H21/02 ,  C07H21/04 ,  C12P21/04

CPC分类号： C12N9/0042 ,  C07K14/40 ,  C12N9/0071 ,  C12N9/0077 ,  C12P7/44

摘要： Novel genes have been isolated which encode cytochrome P450 and NADPH reductase enzymes of the ω-hydroxylase complex of C. tropicalis 20336. Vectors including these genes, transfected host cells and transformed host cells are provided. Methods of producing of cytochrome P450 and NADPH reductase enzymes are also provided which involve transforming a host cell with a gene encoding these enzymes and culturing the cells. Methods of increasing the production of a dicarboxylic acid and methods of increasing production of the aforementioned enzymes are also provided which involve increasing in the host cell the number of genes encoding these enzymes. A method for discriminating members of a gene family by quantifying the expression of genes is also provided.

摘要翻译： 已经分离出编码热带假丝酵母20336的ω-羟化酶复合物的细胞色素P450和NADPH还原酶的新型基因。提供了包括这些基因，转染的宿主细胞和转化的宿主细胞的载体。 还提供了生产细胞色素P450和NADPH还原酶的方法，其涉及用编码这些酶的基因转化宿主细胞并培养细胞。 还提供了增加二羧酸生产的方法和增加上述酶的生产的方法，其涉及在宿主细胞中增加编码这些酶的基因的数量。 还提供了通过定量基因表达来区分基因家族成员的方法。