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DNA Polymerases Having Improved Labeled Nucleotide Incorporation Properties

机译:具有改进的标记核苷酸掺入特性的DNA聚合酶

摘要

The present invention relates to mutant DNA polymerases that exhibit reduced discrimination against labeled nucleotides into polynucleotides. The DNA polymerases of the invention have at least one mutation in the nucleotide label interaction region of the enzyme such the mutation results in reduced discrimination against labeled nucleotides. The nucleotide label interaction regions are located at portions of the O-helix, the K-helix, and the inter O-P helical loop of Taq DNA polymerase or analogous positions in other DNA polymerases.;The invention also provides polynucleotides encoding the mutant DNA polymerases, optionally including expression vectors for mutant polymerase recombinant production, and further optionally including host cells containing the polynucleotides. Numerous methods of using the subject DNA polymerases are provided, including uses for chain termination sequencing and PCR. Kits are provided for synthesizing fluorescently labeled polynucleotides and may include a mutant DNA polymerase of the invention and a fluorescently labeled nucleotide.
机译:本发明涉及突变的DNA聚合酶,其对标记为多核苷酸的核苷酸表现出降低的辨别力。本发明的DNA聚合酶在该酶的核苷酸标记相互作用区域中具有至少一个突变,使得该突变导致对标记核苷酸的辨别力降低。核苷酸标记相互作用区域位于Taq DNA聚合酶的O螺旋,K螺旋和OP间螺旋环的部分或其他DNA聚合酶的类似位置上;本发明还提供了编码突变型DNA聚合酶的多核苷酸,任选地包括用于突变体聚合酶重组生产的表达载体,并且进一步任选地包括包含多核苷酸的宿主细胞。提供了使用主题DNA聚合酶的多种方法,包括链终止测序和PCR的用途。提供了用于合成荧光标记的多核苷酸的试剂盒,该试剂盒可以包括本发明的突变DNA聚合酶和荧光标记的核苷酸。

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