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    Hybridization and mismatch discrimination using oligonucleotides conjugated to minor groove binders
    34.
    发明授权
    Hybridization and mismatch discrimination using oligonucleotides conjugated to minor groove binders 失效
    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：US06884584B2

    公开(公告)日：2005-04-26

    申请号：US10127936

    申请日：2002-04-22

    申请人： Joel Hedgpeth ,  Irina A. Afonina ,  Igor V. Kutyavin ,  Eugeny A. Lukhtanov ,  Evgeniy S. Belousov ,  Rich B. Meyer, Jr.

    发明人： Joel Hedgpeth ,  Irina A. Afonina ,  Igor V. Kutyavin ,  Eugeny A. Lukhtanov ,  Evgeniy S. Belousov ,  Rich B. Meyer, Jr.

    IPC分类号： C12N15/09 ,  C07H19/23 ,  C07H21/00 ,  C07H21/02 ,  C07H21/04 ,  C12Q1/68 ,  G01N21/78 ,  G01N33/483 ,  G01N33/53 ,  G01N33/566 ,  C07H19/04 ,  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： 小沟结合分子如1,2-二氢 - （3H） - 吡咯并[3,2-e]吲哚-7-羧酸酯（CDPI 3 N 3）的三聚体和 寡核苷酸形成具有互补靶序列的异常稳定的杂交体，其中连接的CDPI 3 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。

    Hybridization and mismatch discrimination using oligonucleotides conjugated to minor groove binders
    35.
    发明授权
    Hybridization and mismatch discrimination using oligonucleotides conjugated to minor groove binders 失效
    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：US06312894B1

    公开(公告)日：2001-11-06

    申请号：US09054832

    申请日：1998-04-03

    申请人： Joel Hedgpeth ,  Irina A. Afonina ,  Igor V. Kutyavin ,  Eugeny A. Lukhtanov ,  Evgeniy S. Belousov ,  Rich B. Meyer, Jr.

    发明人： Joel Hedgpeth ,  Irina A. Afonina ,  Igor V. Kutyavin ,  Eugeny A. Lukhtanov ,  Evgeniy S. Belousov ,  Rich B. Meyer, Jr.

    IPC分类号： C12Q168

    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。

    Selective binding complementary oligonucleotides
    38.
    发明授权
    Selective binding complementary oligonucleotides 失效
    选择性结合互补寡核苷酸

    公开(公告)号：US5912340A

    公开(公告)日：1999-06-15

    申请号：US539097

    申请日：1995-10-04

    申请人： Igor V. Kutyavin ,  Jinsuk Woo ,  Eugeny A. Lukhtanov ,  Rich B. Meyer, Jr. ,  Howard B. Gamper

    发明人： Igor V. Kutyavin ,  Jinsuk Woo ,  Eugeny A. Lukhtanov ,  Rich B. Meyer, Jr. ,  Howard B. Gamper

    IPC分类号： C12N15/09 ,  C07H21/00 ,  C12Q1/68 ,  C07H21/04

    CPC分类号： C07H21/00 ,  C12Q1/68

    摘要： In a matched pair of oligonucleotides (ODNS) each member of the pair is complementary or substantially complementary in the Watson Crick sense to a target sequence of duplex nucleic acid where the two strands of the target sequence are themselves complementary to one another. The ODNs include modified bases of such nature that the modified base forms a stable hydrogen bonded base pair with the natural partner base, but does not form a stable hydrogen bonded base pair with its modified partner. This is accomplished when in a hybridized structure the modified base is capable of forming two or more hydrogen bonds with its natural complementary base, but only one hydrogen bond with its modified partner. Due to the lack of stable hydrogen bonding with each other, the matched pair of oligonucleotides have a melting temperature under physiological or substantially physiological conditions of approximately 40.degree. C. or less. However each of the matched ODN pair of the invention forms a substantially stable hybrid with the target sequence in each strand of the duplex nucleic acid. The hybrids of target duplex nucleic acids formed with the ODN pairs of the invention are useful for gene mapping and in diagnostic and therapeutic applications.

    摘要翻译： 在匹配的一对寡核苷酸（ODNS）中，该对中的每个成员在沃森克里克意义上与双链体核酸的靶序列互补或基本互补，其中靶序列的两条链本身彼此互补。 ODN包括这样的性质的修饰碱基，使得修饰的碱基与​​天然配偶体碱基形成稳定的氢键合碱基对，但不与其修饰的配偶体形成稳定的氢键合碱基对。 这是在杂交结构中完成的，修饰的碱基能够与其天然互补碱形成两个或多个氢键，但与其修饰的配偶体仅形成一个氢键。 由于相互之间缺乏稳定的氢键，匹配的寡核苷酸对在约40℃或更低的生理或基本生理条件下具有熔融温度。 然而，本发明的每个匹配的ODN对与双链体核酸的每条链中的靶序列形成基本稳定的杂交体。 用本发明的ODN对形成的靶双链核酸的杂交体可用于基因作图和诊断和治疗应用。