DNA polymerases with improved activity
AssigneeRoche Molecular Systems, Inc.
InventorsKeith Bauer, Thomas W. Myers, Shawn Suko
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Abstract
Disclosed are DNA polymerases having increased reverse transcriptase efficiency relative to a corresponding, unmodified polymerase. The polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the DNA polymerases.
Claims21
A method for conducting primer extension, comprising:
- contacting a DNA polymerase with a primer, a polynucleotide template, and nucleoside triphosphates under conditions suitable for extension of the primer, thereby producing an extended primer,
- wherein the amino acid of the DNA polymerase corresponding to position 616 of SEQ ID NO:1 is any amino acid other than I, and
- wherein the DNA polymerase has at least 93% amino acid sequence identity to a polymerase selected from the group consisting of SEQ ID NO:1, 3, 4, 5, 6, and 7.
A method for conducting a Mg2+-based reverse transcription polymerase chain reaction (RT-PCR), comprising contacting a DNA polymerase with a primer, a polynucleotide template, Mg2+, and nucleoside triphosphates under conditions suitable for RT-PCR, wherein the DNA polymerase has at least 95% sequence identity to the amino acid sequence of SEQ ID NO:1, 2, 3, 4, 5, 6, or 7 and the amino acid of the DNA polymerase corresponding to position 616 of SEQ ID NO:1 is any amino acid other than I, the amino acid corresponding to position 580 of SEQ ID NO:1 is any amino acid other than D, and the amino acid corresponding to position 709 of SEQ ID NO:1 is any amino acid other than I.
Dependencies are inferred from the claim text.