Human DNA topoisomerase I &agr;
    4.
    发明授权
    Human DNA topoisomerase I &agr; 有权
    人类DNA拓扑异构酶Iα

    公开(公告)号:US06255077B1

    公开(公告)日:2001-07-03

    申请号:US09325430

    申请日:1999-06-04

    Abstract: Disclosed is a human is a hTopI-&agr; polypeptide and DNA (RNA) encoding such hTopI-&agr; polypeptide. Also provided is a procedure for producing such polypeptide by recombinant techniques and antibodies and antagonists against such polypeptide. Also provided are methods of using the antibodies and antagonist inhibitors to inhibit the action of hTopI-&agr; for therapeutic purposes such as an antitumor agent, to detect an autoimmune disease, or retroviral infections and to treat adenocarcinoma of the colon. Diagnostic methods for detecting mutations in the coding sequence and alterations in the concentration of the polypeptides in a sample derived from a host are also disclosed.

    Abstract translation: 公开了人是hTopI-α多肽和编码这种hTopI-α多肽的DNA(RNA)。 还提供了通过重组技术产生这种多肽的方法以及针对这种多肽的抗体和拮抗剂。 还提供了使用抗体和拮抗剂抑制剂抑制hTopI-α用于治疗目的如抗肿瘤剂的作用以检测自身免疫性疾病或逆转录病毒感染并治疗结肠腺癌的方法。 还公开了用于检测编码序列中的突变和衍生自宿主的样品中多肽浓度变化的诊断方法。

    Covalent joining of DNA strands to RNA strands catalyzed by vaccinia topoisomerase
    9.
    发明申请
    Covalent joining of DNA strands to RNA strands catalyzed by vaccinia topoisomerase 有权
    将DNA链与牛痘拓扑异构酶催化的RNA链共价连接

    公开(公告)号:US20100317064A1

    公开(公告)日:2010-12-16

    申请号:US12658764

    申请日:2010-02-12

    Abstract: The present invention provides a method of covalently joining a DNA strand to an RNA strand comprising (a) forming a topoisomerase-DNA intermediate by incubating a DNA cleavage substrate comprising a topoisomerase cleavage site with a topoisomerase specific for that site, wherein the topoisomerase-DNA intermediate has one or more 5′ single-strand tails; and (b) adding to the topoisomerase-DNA intermediate an acceptor RNA strand complementary to the 5′ single-strand tail under conditions permitting a ligation of the covalently bound DNA strand of the topoisomerase-DNA intermediate to the RNA acceptor strand and dissociation of the topoisomerase, thereby covalently joining the DNA strand to the RNA strand. The present invention also provides a method of tagging a 5′ end of an RNA molecule. The present invention further provides a DNA-RNA molecule which has been joined in vitro by the use of a topoisomerase. The present invention also provides a method of tagging a 5′ end of an mRNA. The present invention provides a method of isolating and cloning full-length gene sequences using capped mRNA after subtraction of non-capped RNA.

    Abstract translation: 本发明提供了将DNA链与RNA链共价连接的方法,其包括(a)通过将包含拓扑异构酶切割位点的DNA切割底物与该位点特异性的拓扑异构酶一起孵育而形成拓扑异构酶-DNA中间体,其中拓扑异构酶-DNA 中间体具有一个或多个5'单链尾部; 和(b)在允许拓扑异构酶-DNA中间体与RNA受体链的共价结合的DNA链连接的条件下,向拓扑异构酶-DNA中间体添加与5'单链尾部互补的受体RNA链,并解离 从而将DNA链共价连接到RNA链上。 本发明还提供了标记RNA分子的5'末端的方法。 本发明还提供了通过使用拓扑异构酶体外结合的DNA-RNA分子。 本发明还提供了标记mRNA的5'末端的方法。 本发明提供了在减去未加帽的RNA之后使用加帽的mRNA分离和克隆全长基因序列的方法。

    Methods and compositions for generating recombinant nucleic acid molecules
    10.
    发明申请
    Methods and compositions for generating recombinant nucleic acid molecules 有权
    用于产生重组核酸分子的方法和组合物

    公开(公告)号:US20090326208A1

    公开(公告)日:2009-12-31

    申请号:US12112649

    申请日:2008-04-30

    Abstract: A method of generating a double stranded (ds) recombinant nucleic acid molecule covalently linked in both strands by contacting two or more ds nucleotide sequences with a topoisomerase under conditions such that both termini of at least one end of a first ds nucleotide sequence are covalently linked by the topoisomerase to both termini of at least one end of a second ds nucleotide sequence is provided. Also provided is a method for generating a ds recombinant nucleic acid molecule covalently linked in one strand, by contacting two or more ds nucleotide sequences with a type IA topoisomerase under conditions such that one strand, but not both strands, of one or both ends of a first ds nucleotide sequence are covalently linked by the topoisomerase. Compositions for performing such methods, and compositions generated from such methods also are provided, as are kits containing components useful for conveniently practicing the methods.

    Abstract translation: 通过使两个或多个ds核苷酸序列与拓扑异构酶接触的条件使得在第一个ds核苷酸序列的至少一个末端的两个末端共价连接的情况下,产生双链(ds)重组核酸分子共价连接的双链(ds)重组核酸分子的方法 提供了拓扑异构酶至第二个ds核苷酸序列的至少一个末端的两个末端。 还提供了一种用于产生共价连接在一条链中的ds重组核酸分子的方法,其通过使两个或更多个ds核苷酸序列与IA型拓扑异构酶接触,使得一个或两个末端的一条链但不是两条链 第一个ds核苷酸序列由拓扑异构酶共价连接。 还提供了用于进行这些方法的组合物,以及由这些方法产生的组合物,以及包含用于方便地实施该方法的组分的试剂盒。

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