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    Preparation of polypeptides having an amide carboxyl terminal end
    4.
    发明授权
    Preparation of polypeptides having an amide carboxyl terminal end 失效
    制备具有酰胺羧基末端的多肽

    公开(公告)号：US5457033A

    公开(公告)日：1995-10-10

    申请号：US205816

    申请日：1994-03-04

    申请人： Joachim Engels ,  Wolfgang Konig ,  Hubert Mullner ,  Eugen Uhlmann ,  Waldemar Wetekam

    发明人： Joachim Engels ,  Wolfgang Konig ,  Hubert Mullner ,  Eugen Uhlmann ,  Waldemar Wetekam

    IPC分类号： C12N15/09 ,  C07H21/00 ,  C07H21/04 ,  C07K1/00 ,  C07K7/08 ,  C07K14/00 ,  C07K14/575 ,  C07K14/60 ,  C07K19/00 ,  C12N1/20 ,  C12N15/00 ,  C12N15/16 ,  C12N15/31 ,  C12P21/00 ,  C12P21/02 ,  C12R1/19 ,  C12N15/62 ,  C12N15/70 ,  C12P21/06

    CPC分类号： C07H21/00 ,  C07K14/60 ,  C12P21/02 ,  C07K2319/00 ,  C07K2319/75

    摘要： Polypeptides having a carboxamide carboxyl terminal end and a methionine residue, which is optionally bonded to a bacterial protein, on the amino end can be prepared by synthesis by genetic engineering methods of the corresponding polypeptide having at the C terminal end a glycine residue, and conversion of the latter enzymatically into the amino group of the desired carboxamide group. Peptides which have the aminoacid sequence of growth hormone releasing factor, part sequences thereof, or modifications of these peptides, are readily accessible by this means. The synthesis by genetic engineering methods is advantageously carried out via two gene fragments which are synthesized chemically from smaller, single-stranded structural units. The two gene fragments are then linked enzymatically to give the complete gene, which is incorporated into a suitable vector, amplified there, and the peptide is isolated directly or as a fused protein, and is converted enzymatically into the desired amide.

    摘要翻译： 具有羧酰胺羧基末端的多肽和任选地与细菌蛋白结合的氨基末端的甲硫氨酸残基可以通过在C末端具有甘氨酸残基的相应多肽的遗传工程方法合成，并且转化 后者酶促进入所需羧酰胺基团的氨基。 具有生长激素释放因子的氨基酸序列，其部分序列或这些肽的修饰的肽通过这种方式是容易获得的。 通过基因工程方法的合成有利地通过两个基因片段进行，这两个基因片段由较小的单链结构单元化学合成。 然后将两个基因片段酶促连接以产生完整的基因，其被并入合适的载体中，在那里扩增，并且肽直接或作为融合蛋白分离，并且被酶促转化为所需的酰胺。

    3'(2')-amino- or thiol-modified, fluorescent dye-coupled nucleosides, nucleotides and oligonucleotides, and a process for the preparation thereof
    7.
    发明授权
    3'(2')-amino- or thiol-modified, fluorescent dye-coupled nucleosides, nucleotides and oligonucleotides, and a process for the preparation thereof 失效
    3'（2'） - 氨基或硫醇修饰的荧光染料偶联核苷，核苷酸和寡核苷酸，及其制备方法

    公开(公告)号：US5679785A

    公开(公告)日：1997-10-21

    申请号：US184759

    申请日：1994-01-18

    申请人： Joachim Engels ,  Mathias Herrlein ,  Renate Konrad ,  Matthias Mag

    发明人： Joachim Engels ,  Mathias Herrlein ,  Renate Konrad ,  Matthias Mag

    IPC分类号： C07H21/02 ,  A61K31/7052 ,  A61K31/7088 ,  C07H19/20 ,  C07H21/00 ,  C07H21/04 ,  C12Q1/68 ,  G01N33/52 ,  G01N33/58 ,  C07H19/06 ,  C07H19/16

    CPC分类号： C07H21/00 ,  C12Q1/6813 ,  C12Q1/6816 ,  Y02P20/55

    摘要： 3'-(2')-Amino- or thiol-modified, fluorescent dye-coupled nucleosides, nucleotides and oligonucleotides, and a process for the preparation and the use thereof The OH group located in the 3' and/or 2' position of a nucleoside, nucleotide or oligonucleotide is derivatized to an amino or thiol group and subsequently a fluorescent dye is coupled thereto. The resulting 3'- and/or 2'-amino- and thiol-modified nucleosides, nucleotides and oligonucleotides can then be used for the synthesis of complementary strands in the presence of a template strand or of oligonucleotides and for the detection of genetic material. They have the advantage that the fluorescent label need no longer be attached to the 5' end of the oligonucleotide or to the nucleobase, and thus need not be introduced during the chemical synthesis as in labeling techniques hitherto known, while the known and conventional methods have the disadvantage that only a few polymerases can be employed for the synthesis, the acceptance of the triphosphates by the polymerases diminishes and, moreover, a large substrate excess is necessary.

    摘要翻译： 3' - （2'） - 氨基或硫醇修饰的荧光染料偶联的核苷，核苷酸和寡核苷酸，以及其制备和使用的方法。位于3'和/或2'位置的OH基 将核苷，核苷酸或寡核苷酸衍生为氨基或硫醇基，随后将荧光染料偶联至其上。 然后，所得到的3'-和/或2'-氨基 - 和2'-氨基 - 和巯基修饰的核苷，核苷酸和寡核苷酸可用于在模板链或寡核苷酸存在下合成互补链，并用于遗传物质的检测。 它们具有的优点是荧光标记不再需要连接到寡核苷酸的5'末端或核碱基，因此在化学合成过程中不需要像过去已知的标记技术那样引入荧光标记，而已知和常规方法具有 缺点是只有少数聚合酶可用于合成，聚合酶对三磷酸酯的接受性降低，此外，需要大量的底物过量。

    3'-(2')-amimo- or thiol-modified, fluorescent dye-coupled nucleosides, nucleotides and oligonucleotides
    8.
    发明授权
    3'-(2')-amimo- or thiol-modified, fluorescent dye-coupled nucleosides, nucleotides and oligonucleotides 失效
    3' - （2'） - 氨基或硫醇修饰的荧光染料偶联核苷，核苷酸和寡核苷酸

    公开(公告)号：US5668269A

    公开(公告)日：1997-09-16

    申请号：US456734

    申请日：1995-06-01

    申请人： Joachim Engels ,  Mathias Herrlein ,  Renate Konrad ,  Matthias Mag

    发明人： Joachim Engels ,  Mathias Herrlein ,  Renate Konrad ,  Matthias Mag

    IPC分类号： C07H21/02 ,  A61K31/7052 ,  A61K31/7088 ,  C07H19/20 ,  C07H21/00 ,  C07H21/04 ,  C12Q1/68 ,  G01N33/52 ,  G01N33/58 ,  C07H19/06 ,  C07H19/16

    CPC分类号： C07H21/00 ,  C12Q1/6813 ,  C12Q1/6816 ,  Y02P20/55

    摘要： 3'-(2')-Amino- or thiol-modified, fluorescent dye-coupled nucleosides, nucleotides and oligonucleotides, and a process for the preparation and the use thereof.The OH group located in the 3' and/or 2' position of a nucleoside, nucleotide or oligonucleotide is derivatized to an amino or thiol group and subsequently a fluorescent dye is coupled thereto. The resulting 3'- and/or 2'-amino- and thiol-modified nucleosides, nucleotides and oligonucleotides can then be used for the synthesis of complementary strands in the presence of a template strand or of oligonucleotides and for the detection of genetic material. They have the advantage that the fluorescent label need no longer be attached to the 5' end of the oligonucleotide or to the nucleobase, and thus need not be introduced during the chemical synthesis as in labeling techniques hitherto known, while the known and conventional methods have the disadvantage that only a few polymerases can be employed for the synthesis, the acceptance of the triphosphates by the polymerases diminishes and, moreover, a large substrate excess is necessary.

    摘要翻译： 3' - （2'） - 氨基或硫醇修饰的荧光染料偶联的核苷，核苷酸和寡核苷酸，以及其制备和使用的方法。 位于核苷，核苷酸或寡核苷酸的3'和/或2'位置的OH基团衍生为氨基或硫醇基团，随后将荧光染料偶联至其上。 然后，所得到的3'-和/或2'-氨基 - 和2'-氨基 - 和巯基修饰的核苷，核苷酸和寡核苷酸可用于在模板链或寡核苷酸存在下合成互补链，并用于遗传物质的检测。 它们具有的优点是荧光标记不再需要连接到寡核苷酸的5'末端或核碱基，因此在化学合成过程中不需要像过去已知的标记技术那样引入荧光标记，而已知和常规方法具有 缺点是只有少数聚合酶可用于合成，聚合酶对三磷酸酯的接受性降低，此外，需要大量的底物过量。

    Process for the preparation of nucleoside alkyl-aralkyl- and aryl-phosphonites and -phosphonates
    10.
    发明授权
    Process for the preparation of nucleoside alkyl-aralkyl- and aryl-phosphonites and -phosphonates 失效
    核苷烷基 - 芳烷基 - 和芳基 - 亚膦酸酯和膦酸酯的制备方法

    公开(公告)号：US5166330A

    公开(公告)日：1992-11-24

    申请号：US139112

    申请日：1987-12-28

    申请人： Joachim Engels ,  Alfred Jager

    发明人： Joachim Engels ,  Alfred Jager

    IPC分类号： C07F9/48 ,  C07H19/04 ,  C07H21/00

    CPC分类号： C07H19/04 ,  C07F9/4883 ,  C07H21/00

    摘要： Deoxyribonucleoside phosphonates, thiophosphonates and selenophosphonates are obtained by condensation of a difunctional phosphonylating reagent of the formulaR--PXY,in which R is an inert non-cytotoxic organic radical, X is chlorine or Y and Y is a secondary amino group, with a deoxyribonucleoside of which the 5-hydroxyl group and any exo-amino group presents in the base radical are protected, and further condensation with a nucleoside of which the 3-hydroxyl group and any exo-amino group present in the base radical are protected, and then oxidation. The thiophosphonates and selenophosphonates and the intermediates of the first condensation stage are new.

    摘要翻译： 脱氧核糖核苷酸膦酸酯，硫代膦酸酯和硒膦酸酯通过式R-PXY的二官能膦酰化试剂缩合得到，其中R是惰性非细胞毒性有机基团，X是氯或Y，Y是仲氨基，与脱氧核糖核苷 其中5-羟基和任何外基氨基存在于碱基中的基团被保护，并且与其中3-羟基和任何外基氨基存在于碱基中的核苷进一步缩合得到保护，然后 氧化。 硫代膦酸酯和硒膦酸酯和第一个缩合阶段的中间体是新的。