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    Hybrid polypeptide comparing a biotinylated avidin binding polypeptide fused to a polypeptide of interest
    1.
    发明授权
    Hybrid polypeptide comparing a biotinylated avidin binding polypeptide fused to a polypeptide of interest 失效
    杂交多肽比较与感兴趣的多肽融合的生物素化的抗生物素蛋白结合多肽

    公开(公告)号：US6072039A

    公开(公告)日：2000-06-06

    申请号：US687819

    申请日：1991-04-19

    Applicant: Ferdinand Carl Haase ,  Dean Ervin Cress

    Inventor： Ferdinand Carl Haase ,  Dean Ervin Cress

    IPC: A61K38/00 ,  A61K39/00 ,  A61P35/00 ,  C07K1/22 ,  C07K7/04 ,  C07K14/00 ,  C07K14/005 ,  C07K14/195 ,  C07K14/675 ,  C07K19/00 ,  C12N9/10 ,  C12N15/09 ,  C12N15/62 ,  C12P21/02 ,  C12R1/19 ,  C12R1/91 ,  G01N33/53 ,  G01N33/574 ,  C12N15/00

    CPC classification number: C12N15/62 ,  C07K14/675 ,  C12N9/1018 ,  C07K2319/00 ,  C07K2319/75

    Abstract: A hybrid polypeptide is disclosed, comprising a polypeptide for attachment of a prosthetic group to avidin fused to at least one polypeptide of interest, and a method of making the same. The hybrid polypeptide is produced by recombinant DNA techniques, using a DNA expression vector composed of a DNA fragment coding for the polypeptide for attachment to avidin fused to DNA coding for one or more polypeptides of interest. The hybrid polypeptide may also contain linking amino acid sequences for cleavage of the polypeptide of interest from the polypeptide for attachment using an appropriate proteolytic or chemical reagent. The hybrid polypeptide is expressed in appropriate host cells transformed with the DNA expression vector encoding the hybrid polypeptide, and may be recovered from crude cell extracts in high yield and high purity using avidin affinity chromatography. Following avidin affinity purification, the polypeptide for attachment and polypeptide of interest may be cleaved to yield polypeptide of interest in a highly pure and highly active state.

    Abstract translation: 公开了一种杂合多肽，其包含用于将假基团连接至与至少一种目标多肽融合的抗生物素蛋白的多肽及其制备方法。 通过重组DNA技术，使用由编码多肽的DNA片段组成的DNA表达载体来产生杂交多肽，所述DNA片段与用于编码一种或多种感兴趣多肽的DNA融合的抗生物素蛋白连接。 杂合多肽还可以含有用于使用合适的蛋白水解或化学试剂从所述多肽切割感兴趣多肽的连接氨基酸序列用于连接。 杂交多肽在用编码杂合多肽的DNA表达载体转化的合适的宿主细胞中表达，并且可以使用亲和素亲和层析以高产率和高纯度从粗细胞提取物中回收。 在亲和素亲和纯化之后，可以切割用于附着的多肽和目标多肽，以在高纯度和高活性状态下产生感兴趣的多肽。

    Method for extracellular production of target proteins employing OmpF in E. coli
    4.
    发明授权
    Method for extracellular production of target proteins employing OmpF in E. coli 失效
    使用OmpF在大肠杆菌中细胞外产生靶蛋白的方法

    公开(公告)号：US07491528B2

    公开(公告)日：2009-02-17

    申请号：US10600145

    申请日：2003-06-19

    Applicant: Sang-Yup Lee ,  Ki-Jun Jeong

    Inventor： Sang-Yup Lee ,  Ki-Jun Jeong

    IPC: C12P21/06 ,  C12P21/04 ,  C12N15/64 ,  C12N1/20 ,  C12N15/00 ,  C12N15/09 ,  C12N15/63 ,  C12N15/70 ,  C12N15/74

    CPC classification number: C12P21/02 ,  C07K14/245 ,  C07K14/675 ,  C07K2319/02 ,  C07K2319/035 ,  C07K2319/75 ,  C12N15/625 ,  C12N15/70

    Abstract: The present invention provides an expression vector comprising genes encoding OmpF of E. coli and a desired protein, E.coli transformed with the expression vector, and a method for extracellular production of desired proteins by employing the same. The recombinant expression vector of the invention comprises an ampicillin-resistance gene, the OmpF promoter and the OmpF gene. In accordance with the invention, a desired protein can be produced extracellularly by a simpler method than conventional methods such that: secretory production of OmpF fusion protein begins simultaneously with growth of the cells through constitutive expression employing an OmpF promoter, and as the concentration of cells increases, the amount of secretory production of the protein also increases continuously. Therefore, desired proteins can be produced in large quantities by a high concentration culture of cells.

    Abstract translation: 本发明提供了包含编码大肠杆菌OmpF和所需蛋白质的基因的表达载体，用表达载体转化的大肠杆菌，以及通过使用该方法细胞外产生所需蛋白质的方法。 本发明的重组表达载体包含氨苄青霉素抗性基因，OmpF启动子和OmpF基因。 根据本发明，可以通过比常规方法更简单的方法在细胞外产生所需的蛋白质，使得OmpF融合蛋白的分泌生成与通过使用OmpF启动子的组成型表达的细胞的生长同时开始，并且作为细胞的浓度 增加，蛋白质的分泌生产量也不断增加。 因此，可以通过高浓度细胞培养物大量生产所需的蛋白质。

    Carboxyl terminus analogs of .beta.-endorphin
    6.
    发明授权
    Carboxyl terminus analogs of .beta.-endorphin 失效
    β-内啡肽的羧基末端类似物

    公开(公告)号：US4250087A

    公开(公告)日：1981-02-10

    申请号：US102094

    申请日：1979-12-10

    Applicant: Choh H. Li

    Inventor： Choh H. Li

    IPC: A61K38/00 ,  C07K14/675 ,  C07C103/52

    CPC classification number: C07K14/675 ,  A61K38/00 ,  Y10S514/809

    Abstract: Novel carboxyl terminus analogs of .beta.-endorphin are described. The carboxyl terminus of such analogs is selected from lower alkyl amide, glycine, glycinamide, or polyglycine. These analogs exhibit either greater analgesic activity than the parent .beta.-endorphin and/or increased binding activity in an opiate binding assay.

    Abstract translation: 描述了β-内啡肽的新型羧基末端类似物。 这种类似物的羧基末端选自低级烷基酰胺，甘氨酸，甘氨酰胺或聚甘氨酸。 这些类似物在阿片剂结合测定中表现出比母体β-内啡肽更大的镇痛活性和/或增加的结合活性。

    Manufacture and expression of structural genes
    10.
    发明授权
    Manufacture and expression of structural genes 失效
    结构基因的制造和表达

    公开(公告)号：US4652639A

    公开(公告)日：1987-03-24

    申请号：US375493

    申请日：1982-05-06

    Applicant: Yitzhak Stabinsky

    Inventor： Yitzhak Stabinsky

    IPC: G01N33/50 ,  C07H1/00 ,  C07H21/00 ,  C07H21/04 ,  C07K14/00 ,  C07K14/575 ,  C07K14/655 ,  C07K14/675 ,  C07K16/00 ,  C07K19/00 ,  C12N15/00 ,  C12N15/09 ,  C12N15/10 ,  C12P21/00 ,  C12R1/19 ,  G01N33/60 ,  C07H21/02 ,  C12N1/00 ,  C12P19/34 ,  C12P21/02

    CPC classification number: C07H21/00 ,  C07K14/675 ,  C12N15/10 ,  G01N33/60

    Abstract: Described are rapid and highly efficient procedures for the total synthesis of linear, double stranded DNA sequences of up to about 200 base pairs, which sequences may comprise entire structural genes. Illustratively disclosed is the preparation and expression of manufactured genes, including fusion genes, capable of directing synthesis of human .beta.-endorphin and of proteins which differ from human .beta.-endorphin in terms of the identity or relative position of one or more amino acids. Manufactured genes preferably include codons selected from among alternative codons specifying the same amino acid on the basis of preferential expression in a projected host microorganism (e.g., E. coli) to be transformed.

    Abstract translation: 描述了用于全合成高达约200个碱基对的线性双链DNA序列的快速和高效的方法，该序列可以包含整个结构基因。 示例性地公开了在一个或多个氨基酸的同一性或相对位置方面，制备和表达能够引导人β-内啡肽和不同于人β-内啡肽的蛋白质的合成基因，包括融合基因。 制造的基因优选包括基于待转化的投影宿主微生物（例如大肠杆菌）中优先表达的选自指定相同氨基酸的替代密码子的密码子。

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