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    Hybridization and mismatch discrimination using oligonucleotides conjugated to minor groove binders
    4.
    发明申请
    Hybridization and mismatch discrimination using oligonucleotides conjugated to minor groove binders 审中-公开
    使用与小沟结合物缀合的寡核苷酸的杂交和错配鉴别

    公开(公告)号：US20050064463A1

    公开(公告)日：2005-03-24

    申请号：US10870578

    申请日：2004-06-16

    申请人： Joel Hedgpeth ,  Irina Afonina ,  Igor Kutyavin ,  Eugeny Lukhtanov ,  Evgeniy Belousov ,  Rich Meyer

    发明人： Joel Hedgpeth ,  Irina Afonina ,  Igor Kutyavin ,  Eugeny Lukhtanov ,  Evgeniy Belousov ,  Rich Meyer

    IPC分类号： C12N15/09 ,  C07H19/23 ,  C07H21/00 ,  C07H21/02 ,  C07H21/04 ,  C12Q1/68 ,  C12Q1/6813 ,  G01N21/78 ,  G01N33/483 ,  G01N33/53 ,  G01N33/566 ,  C12P19/34

    CPC分类号： C07H21/00 ,  C07H19/23 ,  C12Q1/6813 ,  C12Q2527/107 ,  C12Q2525/197

    摘要： Conjugates between a minor groove binding molecule, such as the trimer of 1,2-dihydro-(3H)-pyrrolo[3,2-e]indole-7-carboxylate (CDPI3), and an oligonucleotide form unusually stable hybrids with complementary target sequences, in which the tethered CDPI3 group resides in the minor groove of the duplex. These conjugates can be used as probes and primers. Due to their unusually high binding affinity, conjugates as short as 8-mers can be used as amplification primers with high specificity and efficiency. MGB conjugation also increases the discriminatory power of short oligonucleotides, providing enhanced detection of nucleotide sequence mismatches by short oligonucleotides. The MGB-conjugated probes and primers described herein facilitate various analytic and diagnostic procedures, such as amplification reactions, PCR, detection of single-nucleotide polymorphisms, gene hunting, differential display, fluorescence energy transfer, hydrolyzable probe assays and others; by allowing the use of shorter oligonucleotides, which have higher specificity and better discriminatory power.

    摘要翻译： 小沟结合分子（例如1,2-二氢 - （3H） - 吡咯并[3,2-e]吲哚-7-羧酸酯（CDPI3）的三聚体和寡核苷酸之间的缀合物形成非常稳定的具有互补靶的杂交体 序列，其中系留的CDPI3基团位于双链体的小沟槽中。 这些缀合物可以用作探针和引物。 由于其非常高的结合亲和力，短至8聚体的缀合物可以用作高特异性和高效的扩增引物。 MGB缀合还增加了短寡核苷酸的鉴别能力，提供了通过短寡核苷酸增强核苷酸序列错配检测。 本文所述的MGB缀合的探针和引物有助于各种分析和诊断程序，如扩增反应，PCR，单核苷酸多态性检测，基因敲除，差异显示，荧光能量转移，水解性探针测定等。 通过允许使用具有更高特异性和更好的辨别力的较短的寡核苷酸。