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    • 65. 发明授权
    • Mutant T7 polymerases
    • 突变T7聚合酶
    • US09062292B2
    • 2015-06-23
    • US12807751
    • 2010-09-13
    • Jack Coleman
    • Jack Coleman
    • C12N9/12
    • C12N9/1247
    • Provided are mutant polymerases that comprise a deletion of at least four amino acids among the amino acids at positions corresponding to 167-174 of SEQ ID NO:1. Also provided are mutant polymerases having greater resistance to 30 mM NaCl, 7.5 mM phosphate, or 20 μg/ml single stranded DNA than a wild-type T7 RNA polymerase having SEQ ID NO:1 or a wild-type T3 RNA polymerase having SEQ ID NO:3. Nucleic acids comprising a nucleotide sequence encoding any of the above mutant polymerases are also provided, as are vectors comprising those nucleic acids and host cells transformed with the vectors Additionally, methods of amplifying mRNA using the mutant polymerases described herein are also provided. Further, compositions comprising any of the mutant polymerases described herein, and a reagent at a concentration that is inhibitory to wild-type T7 RNA polymerase is provided.
    • 提供的突变体聚合酶包含在对应于SEQ ID NO:1的167-174位置的氨基酸中至少4个氨基酸的缺失。 还提供了比具有SEQ ID NO:1的野生型T7RNA聚合酶或具有SEQ ID NO:1的野生型T3 RNA聚合酶对30mM NaCl,7.5mM磷酸酯或20μg/ ml单链DNA具有更大抗性的突变型聚合酶 NO:3。 还提供了包含编码任何上述突变型聚合酶的核苷酸序列的核酸,以及包含用载体转化的那些核酸和宿主细胞的载体。另外,还提供了使用本文所述的突变型聚合酶扩增mRNA的方法。 此外,提供了包含本文所述的任何突变型聚合酶的组合物和对野生型T7RNA聚合酶具有抑制浓度的试剂。