公开(公告)号：US06221630B1

公开(公告)日：2001-04-24

申请号：US09275680

申请日：1999-03-24

申请人： James E. Hopper

发明人： James E. Hopper

IPC分类号： C12P2102

CPC分类号： C12N15/81

摘要： The invention provides methods and recombinant expression constructs for inducing and sustaining high-level production of a recombinant polypeptide in yeast. The invention specifically provides high copy number recombinant expression constructs that express high levels of trans-acting transcription factors that in turn induce expression of a recombinant nucleic acid encoding a heterologous or endogenous recombinant polypeptide. The invention more specifically provides constructs that express galactose-inducible and temperature-sensitive transcription factors. The invention also provides constructs comprising nucleic acids the transcription of which is regulated by the transcription factors expressed by the construct. The invention also provides yeast cells transformed by the recombinant expression constructs of the invention that permit sustained high-level expression of a recombinant polypeptide. The invention further provides methods for producing yeast cells transformed with the recombinant expression constructs of the invention, and methods for inducing high-level expression of a recombinant polypeptide in said transformed yeast cells. The invention also provides such recombinant polypeptides produced by the methods and the transformed yeast cells of the invention.

摘要翻译： 本发明提供用于诱导和维持酵母中重组多肽的高水平产生的方法和重组表达构建体。 本发明特别提供了表达高水平的反式作用转录因子的高拷贝数重组表达构建体，其又诱导编码异源或内源重组多肽的重组核酸的表达。 本发明更具体地提供表达半乳糖诱导型和温度敏感性转录因子的构建体。 本发明还提供了包含其转录由构建体表达的转录因子调节的核酸的构建体。 本发明还提供了通过本发明的重组表达构建体转化的允许持续高水平表达重组多肽的酵母细胞。 本发明进一步提供用于产生用本发明的重组表达构建体转化的酵母细胞的方法，以及用于在所述转化的酵母细胞中诱导重组多肽的高水平表达的方法。 本发明还提供了通过本发明的方法和转化的酵母细胞产生的这种重组多肽。

公开(公告)号：US06183985B2

公开(公告)日：2001-02-06

申请号：US08397553

申请日：1995-03-01

申请人： Jeffrey R. Shuster

发明人： Jeffrey R. Shuster

IPC分类号： C12P2102

CPC分类号： C12N15/90 ,  C07K14/503 ,  C07K2319/00 ,  C07K2319/75 ,  C12N15/81

摘要： A method is provided for expressing non-yeast proteins under the control of ADH2 regulatory sequences in a yeast host having enhanced expression of ADR I. Transformed yeast hosts are also provided.

摘要翻译： 提供了在具有增强的ADR I表达的酵母宿主中在ADH2调节序列控制下表达非酵母蛋白的方法。还提供了转化的酵母宿主。

公开(公告)号：US06180363B2

公开(公告)日：2001-01-30

申请号：US09158767

申请日：1998-09-23

申请人： Yannick Batard ,  Francis Durst ,  Michel Schalk ,  Daniele Werck-Reichhart

发明人： Yannick Batard ,  Francis Durst ,  Michel Schalk ,  Daniele Werck-Reichhart

IPC分类号： C12P2102

CPC分类号： C12N9/0042 ,  C12N15/67 ,  C12N15/81

摘要： The present invention relates to a DNA sequence which encodes a protein of interest which contains regions having a high content of codons which are poorly suited to yeasts, characterized in that a sufficient number of codons which are poorly suited to yeasts is replaced with corresponding codons which are well-suited to yeasts in the said regions having a high content of codons which are poorly suited to yeasts. The present invention relates, more specifically, to DNA sequences which originate from dicotyledonous or monocotyledonous plants, in particular plants of the graminae family which are selected, in particular, from among wheat, barley, oats, rice, maize, sorghum and cane sugar. The present invention also relates to transformed yeasts which contain a DNA sequence according to the invention.

摘要翻译： 本发明涉及编码感兴趣的蛋白质的DNA序列，该DNA序列含有密度较高且不适于酵母的密度较高的区域，其特征在于足够数量的不适于酵母的密码子被相应的密码子替换， 在不适合酵母的密码子含量较高的所述区域中适用于酵母菌。 本发明更具体地涉及源于双子叶植物或单子叶植物的DNA序列，特别是特别是选自小麦，大麦，燕麦，稻，玉米，高粱和蔗糖中的格氏族的植物。 本发明还涉及含有根据本发明的DNA序列的转化酵母。

公开(公告)号：US06800457B2

公开(公告)日：2004-10-05

申请号：US10320176

申请日：2002-12-16

申请人： Kanaka Raju Koduri ,  John T. Miller ,  Pallaiah Thammana

发明人： Kanaka Raju Koduri ,  John T. Miller ,  Pallaiah Thammana

IPC分类号： C12P2102

CPC分类号： C12N15/85 ,  C07K14/70521 ,  C07K2319/00

摘要： A specific locus (hot spot) for recombinant gene expression has been identified in the genome of Chinese hamster ovary cells. A DNA vector containing the hot spot causes high levels of recombinant gene expression following transfection and stable integration. The selection and cloning of the specific locus and the expression of recombinant genes is disclosed, as are the DNA vectors and the host cells.

摘要翻译： 已经在中国仓鼠卵巢细胞的基因组中鉴定了重组基因表达的特异性位点（热点）。 含有热点的DNA载体在转染和稳定整合后导致高水平的重组基因表达。 公开了特异性基因座的选择和克隆以及重组基因的表达，以及DNA载体和宿主细胞。

公开(公告)号：US06750045B2

公开(公告)日：2004-06-15

申请号：US10201168

申请日：2002-07-23

申请人： Bruce L. Zamost

发明人： Bruce L. Zamost

IPC分类号： C12P2102

CPC分类号： C12N1/18 ,  C07K14/755

摘要： A yeast fermentation medium that does not contain yeast cell extract but is comprised of a mixture of oleic acid, lactic acid and palmitic acid. Using this fermentation medium yields of factor XIII were increased 250%.

摘要翻译： 不含酵母细胞提取物但由油酸，乳酸和棕榈酸的混合物组成的酵母发酵培养基。 使用该发酵培养基，因子XIII的产量增加了250％。

公开(公告)号：US06686179B2

公开(公告)日：2004-02-03

申请号：US09984186

申请日：2001-10-29

申请人： Reinhard Fleer ,  Alain Fournier ,  Jean-Dominique Guitton ,  Gérard Jung ,  Patrice Yeh

发明人： Reinhard Fleer ,  Alain Fournier ,  Jean-Dominique Guitton ,  Gérard Jung ,  Patrice Yeh

IPC分类号： C12P2102

CPC分类号： B65B69/00 ,  A61K38/21 ,  A61K38/38 ,  A61K47/642 ,  A61K47/643 ,  C07K14/575 ,  C07K14/755 ,  C07K14/76 ,  C07K14/765 ,  C07K2319/00 ,  C07K2319/02 ,  C07K2319/31 ,  C07K2319/33 ,  C12N9/6456 ,  A61K2300/00

摘要： Biologically active polypeptides comprising a therapeutically active polypeptide fused to human serum albumin or a variant thereof, methods for the preparation thereof, nucleotide sequences encoding such fusion polypeptides, expression cassettes comprising such nucleotide sequences, self-replicating plasmids containing such expression cassettes, and pharmaceutical compositions containing said fusion polypeptides.

摘要翻译： 包含与人血清白蛋白或其变体融合的治疗活性多肽的生物活性多肽，其制备方法，编码这种融合多肽的核苷酸序列，包含该核苷酸序列的表达盒，含有该表达盒的自体复制质粒和药物组合物 含有所述融合多肽。

公开(公告)号：US06670151B1

公开(公告)日：2003-12-30

申请号：US09485639

申请日：2000-04-25

申请人： Manfred Baetscher

发明人： Manfred Baetscher

IPC分类号： C12P2102

CPC分类号： C07K14/475

摘要： This invention relates to newly identified polynucleotides, polypeptides encoded by such polynucleotides, the use of such polynucleotides and polypeptides, as well as the production and isolation of such polynucleotides and polypeptides. More particularly, the polynucleotides and polypeptides of the present invention have been identified as porcine stem cell factors, and in particular membrane-bound porcine stem cell factors, and still more particularly as being involved in the culture of pluripotent or totipotent porcine cells.

摘要翻译： 本发明涉及新鉴定的多核苷酸，由这种多核苷酸编码的多肽，这种多核苷酸和多肽的用途，以及这些多核苷酸和多肽的产生和分离。 更具体地，本发明的多核苷酸和多肽已被鉴定为猪干细胞因子，特别是膜结合的猪干细胞因子，更特别地，涉及多能性或全能猪细胞的培养。

公开(公告)号：US06635451B2

公开(公告)日：2003-10-21

申请号：US09769863

申请日：2001-01-25

申请人： Pradip Mukerji ,  Yung-Sheng Huang ,  Tapas Das ,  Jennifer Thurmond ,  Suzette L. Pereira

发明人： Pradip Mukerji ,  Yung-Sheng Huang ,  Tapas Das ,  Jennifer Thurmond ,  Suzette L. Pereira

IPC分类号： C12P2102

CPC分类号： C12N9/0083 ,  C12N15/8247 ,  C12P7/6427 ,  C12P7/6472

摘要： The subject invention relates to the identification of genes involved in the desaturation of polyunsaturated fatty acids at carbon 5 (i.e., “&Dgr;5-desaturase”) and at carbon 6 (i.e., “&Dgr;6-desaturase”) and to uses thereof. In particular, &Dgr;5-desaturase may be utilized, for example, in the conversion of dihomo-&ggr;-linolenic acid (DGLA) to arachidonic acid (AA) and in the conversion of 20:4n-3 to eicosapentaenoic acid (EPA). Delta-6 desaturase may be used, for example, in the conversion of linoleic (LA) to &ggr;-linolenic acid (GLA). AA or polyunsaturated fatty acids produced therefrom may be added to pharmaceutical compositions, nutritional compositions, animal feeds, as well as other products such as cosmetics.

公开(公告)号：US06610514B2

公开(公告)日：2003-08-26

申请号：US09908855

申请日：2001-07-20

申请人： Keiji Kondo ,  Yutaka Miura

发明人： Keiji Kondo ,  Yutaka Miura

IPC分类号： C12P2102

CPC分类号： C12N9/2411 ,  C07K14/43 ,  C12N9/2408 ,  C12N15/815 ,  C12N15/905 ,  C12Y302/01001

摘要： An object of the present invention is to provide a vector which can be integrated into a yeast chromosome in a high number of copies. Another object of the present invention is to provide a modified vector which can be integrated into the yeast chromosome in a high number of copies and of which expression units stably maintain on the chromosome. The vector according to the present invention comprises a marker gene for selecting transformants, a shortened promoter sequence which is operably linked to the marker gene and a sequence homologous to the chromosomal DNA of Candida utilis, and optionally a heterologous gene or a gene derived from C. utilis, wherein the vector is linearized by cleaving within said homologous DNA sequence or at both ends of the homologous DNA sequence with restriction enzymes, and wherein the heterologous gene or the gene derived from C. utilis can be integrated into the chromosomal DNA of C. utilis by homologous recombination.

公开(公告)号：US06562588B2

公开(公告)日：2003-05-13

申请号：US08871076

申请日：1997-06-09

申请人： Thomas G. Warner ,  Mary B. Sliwkowski

发明人： Thomas G. Warner ,  Mary B. Sliwkowski

IPC分类号： C12P2102

CPC分类号： C12P21/005 ,  A61K38/00 ,  C12N9/2402 ,  C12N15/01 ,  C12Y302/01018

摘要： A recombinant cell line has a constitutive sialidase whose functional expression is disrupted, for example by homologous recombination or using antisense RNA. Sialidase is purified from cell culture fluid of Chinese hamster ovary cells. Nucleic acid encoding sialidase is obtained using an oligonucleotide probe designed using amino acid sequence data on the sialidase.

摘要翻译： 重组细胞系具有组成型唾液酸酶，其功能性表达被破坏，例如通过同源重组或使用反义RNA。 从中国仓鼠卵巢细胞的细胞培养液中纯化唾液酸酶。 使用使用唾液酸酶上的氨基酸序列数据设计的寡核苷酸探针获得核酸编码唾液酸酶。