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    Stabilized expression vectors containing lambda pl promoter and the gene for the CI434 repressor, plasmids containing the vectors, hosts containing the plasmids and related methods
    52.
    发明授权
    Stabilized expression vectors containing lambda pl promoter and the gene for the CI434 repressor, plasmids containing the vectors, hosts containing the plasmids and related methods 失效
    包含λpl启动子和CI434阻遏物的基因的稳定表达载体，含有载体的质粒，含有质粒的宿主和相关方法

    公开(公告)号：US5147789A

    公开(公告)日：1992-09-15

    申请号：US780353

    申请日：1991-10-22

    申请人： Amos B. Oppenheim ,  Hilla Locker-Giladi

    发明人： Amos B. Oppenheim ,  Hilla Locker-Giladi

    IPC分类号： A23K1/165 ,  A61K38/44 ,  C07K14/61 ,  C07K14/775 ,  C12N1/21 ,  C12N9/02 ,  C12N15/69 ,  C12N15/73

    CPC分类号： C12N9/0089 ,  A23K20/168 ,  A61K38/446 ,  C07K14/61 ,  C07K14/775 ,  C12N15/69 ,  C12N15/73

    摘要： An improved vector upon introduction into a suitable host containing the thermolabile repressor C.sub.I renders the host capable of effecting expression of a desired gene. The vector is a double-stranded DNA molecule which includes in 5' to 3' order the following: the promoter and operator P.sub.L O.sub.L from lambda bacteriophage; the N utilization site; a first restriction enzyme site permitting replacement of the ribosomal binding site which follows thereafter; a ribosomal bindig site; an ATG initiation codon or DNA which is converted into an ATG initiation codon upon insertion of the desired gene into the vector; a second restriction enzyme site for inserting the gene in phase with the ATG codon; a T.sub.1 T.sub.2 rRNA transcription termination sequence; an origin of replication; and a fragment designated CI.sup.434 on which is included the gene for the repressor protein and its associated promoter and operator. Additionally, the vector may include a gene associated with a selectable or identifiable phenotypic trait which is manifested when the vector is present in the host. The distance between the 3' end of P.sub.L O.sub.L and the 5' end of the N utilization site is less than about 80 base pairs. The distance between the 3' end of the N utilization site and the 5' end of the ribosomal binding site is less than about 300 base pairs. Plasmids have been constructed from the vectors and used to produce bovine growth hormones.

    摘要翻译： 在引入含有热不稳定阻遏物CI的合适宿主中时的改良载体使宿主能够实现所需基因的表达。 载体是双链DNA分子，其以5'至3'的顺序包括以下：来自λ噬菌体的启动子和操纵子PLOL; N利用场所; 第一限制性内切酶位点，其允许置换随后的核糖体结合位点; 核糖体结合位点; ATG起始密码子或DNA，将所需基因插入载体后转化为ATG起始密码子; 用于将基因插入与ATG密码子同相的第二限制酶切位点; T1T2 rRNA转录终止序列; 复制的起源; 以及指定为CI434的片段，其中包含用于阻遏蛋白的基因及其相关启动子和操纵子。 此外，载体可以包括与可选择或可鉴定的表型性状相关的基因，其在载体存在于宿主中时显示。 PLOL的3'端与N利用位点的5'端之间的距离小于约80个碱基对。 N利用位点的3'末端与核糖体结合位点的5'末端之间的距离小于约300个碱基对。 质粒已经从载体构建并用于产生牛生长激素。

    Plasmids with conditional uncontrolled replication behavior
    56.
    发明授权
    Plasmids with conditional uncontrolled replication behavior 失效
    具有条件不受控制的复制行为的质粒

    公开(公告)号：US4806471A

    公开(公告)日：1989-02-21

    申请号：US610765

    申请日：1984-05-16

    申请人： Soren Molin ,  Janice A. Light ,  Jens E. L. Larsen

    发明人： Soren Molin ,  Janice A. Light ,  Jens E. L. Larsen

    IPC分类号： C12N15/68 ,  C12N15/69 ,  C12N15/00 ,  C12N1/20 ,  C12P19/34 ,  C12P21/00

    CPC分类号： C12N15/68 ,  C12N15/69

    摘要： Plasmids useful as cloning vectors carrying an inserted regulatable promoter, transcription from the promoter regulating plasmid replication. Increased transcription leads to a substantially increased or uncontrolled plasmid copy number when host microorganisms containing the plasmid are cultivated under conditions securing such increased transcription. The regulatable promoter is preferably .lambda. P.sub.R, the activity of which is controlled by a temperature-sensitive .lambda. cl repressor. To obtain gene products of the plasmids, host microorganisms to which such plasmids have been transformed are desirably cultivated at about 30.degree. C. to secure a constant, low plasmid copy number during the seeding and multiplication stages of the microorganisms, after which the temperature is preferably shifted to about 36.degree.-42.degree. C. to obtain an uncontrolled plasmid copy number and an amplified amount of gene product.

    摘要翻译： 质粒用作携带插入的可调节启动子的克隆载体，来自启动子的转录调节质粒复制。 当含有质粒的宿主微生物在确保这种增加的转录的条件下培养时，增加的转录导致显着增加或不受控制的质粒拷贝数。 可调节的启动子优选为λPR，其活性由温度敏感的λ1c阻遏物控制。 为了获得质粒的基因产物，优选在约30℃培养这些质粒转化的宿主微生物，以在微生物的播种和繁殖阶段期间确保恒定的低质粒拷贝数，之后温度为 优选转移到约36℃-42℃以获得不受控制的质粒拷贝数和扩增量的基因产物。

    Vectors suitable for detection of eukaryotic DNA regulatory sequences
    57.
    发明授权
    Vectors suitable for detection of eukaryotic DNA regulatory sequences 失效
    适用于检测真核DNA调控序列的载体

    公开(公告)号：US4761367A

    公开(公告)日：1988-08-02

    申请号：US669224

    申请日：1984-11-07

    申请人： Marshall H. Edgell ,  Steven K. Nordeen ,  Clyde A. Hutchinson, III

    发明人： Marshall H. Edgell ,  Steven K. Nordeen ,  Clyde A. Hutchinson, III

    IPC分类号： C12N15/69 ,  C12N15/85 ,  C12Q1/68 ,  C12N5/00 ,  C12N15/00 ,  C12P21/00

    CPC分类号： C12N15/69 ,  C12N15/85 ,  C12Q1/6897

    摘要： The present invention relates to vectors useful for identifying eukaryotic regulatory sequences and to a method for identifying these sequences. The vectors comprise a first nucleotide sequence which allows for replication in a eukaryotic host, a second nucleotide sequence which codes for a first product that controls replication at said first sequence, and a third nucleotide sequence which codes for a second product that is detectable. The method comprises inserting putative eukaryotic regulatory sequences at a position on the 5'-side of the second sequence and measuring the production of the second product.

    摘要翻译： 本发明涉及用于鉴定真核调节序列的载体和鉴定这些序列的方法。 载体包含允许在真核宿主中复制的第一核苷酸序列，编码控制所述第一序列复制的第一产物的第二核苷酸序列和编码可检测的第二产物的第三核苷酸序列。 该方法包括在第二序列的5'侧的位置插入推定的真核调节序列并测量第二产物的产生。

    Method for producing enzymatically active eucaryotic sod in bacteria
    58.
    发明授权
    Method for producing enzymatically active eucaryotic sod in bacteria 失效
    在细菌中生产酶活性真核细胞的方法

    公开(公告)号：US4742004A

    公开(公告)日：1988-05-03

    申请号：US644105

    申请日：1984-08-27

    申请人： Jacob R. Hartman ,  Dov Kanner ,  Daniel Bartfeld

    发明人： Jacob R. Hartman ,  Dov Kanner ,  Daniel Bartfeld

    IPC分类号： A23K1/165 ,  A61K38/44 ,  C07K14/61 ,  C07K14/775 ,  C12N9/02 ,  C12N15/69 ,  C12N15/73 ,  C12P21/02 ,  C12N15/00

    CPC分类号： C12N9/0089 ,  A23K20/168 ,  A61K38/446 ,  C07K14/61 ,  C07K14/775 ,  C12N15/69 ,  C12N15/73

    摘要： A method of producing enzymatically active eucaryotic superoxide dismutase or an analog thereof in a bacterial cell which contains and is capable of expressing a DNA sequence encoding the superoxide dismutase or analog thereof comprising maintaining the bacterial cell under suitable conditions and in a suitable production medium. The production medium is supplemented with an amount of Cu++ so that the concentration of Cu++ in the medium is greater than about about 2 ppm.The invention also concerns methods of recovering purified enzymatically active eucaryotic superoxide dismutase or analogs from bacterial cells producing the superoxide dismutase or analog.

    摘要翻译： 一种在细菌细胞中生产酶活性的真核超氧化物歧化酶或其类似物的方法，其含有并能够表达编码超氧化物歧化酶或其类似物的DNA序列，其包括在合适的条件下和在合适的生产培养基中保持细菌细胞。 生产培养基补充一定量的Cu ++，使得培养基中Cu ++的浓度大于约2ppm。 本发明还涉及从产生超氧化物歧化酶或类似物的细菌细胞中回收纯化的酶促活性的真核超氧化物歧化酶或类似物的方法。