Mutant Taq polymerase for faster amplification
The invention includes a mutant Taq polymerase, which can significantly extend and amplify a target sequence where the extension conditions are time limited to as little as one second. The mutant Taq polymerase, or a biologically active fragment thereof, has one or more substitutions differing from the wild type as shown in Table I.
1. A mutant Taq polymerase comprising the amino acid mutation E626K (SEQ ID NO: 332) and wherein the remainder of the mutant polymerase is identical to wild type Taq polymerase, as shown in SEQ ID NO: 4 but wherein wild type Taq polymerase does not include the 6-membered histidine tag at its C-terminus and the six immediately preceding Glycine and Serine amino acids shown in SEQ ID NO: 4.
2. The mutant Taq polymerase with the amino acid sequence of SEQ ID NO: 332.
3. A mutant Taq polymerase comprising: one or more of the following amino acid mutations shown in the following even-numbered sequences, and wherein the remainder of the mutant polymerase has at least 80% amino acid sequence identity with wild type Taq polymerase, as shown in SEQ ID NO: 4 but wherein wild type Taq polymerase does not include the 6-membered histidine tag at its C-terminus and the six immediately preceding Glycine and Serine amino acids shown in SEQ ID NO: 4:
G32P (SEQ ID NOS: 402-403), T34I (SEQ ID NOS: 408-409), E39K/E230K (SEQ ID NOS: 28-29), E39K/D320R (SEQ ID NOS: 30-31), E39K/E507K (SEQ ID NOS: 32-33), E39K/E520K (SEQ ID NOS: 34-35), E39K/E537K (SEQ ID NOS: 36-37), E39K/D578R (SEQ ID NOS: 38-39), E39K/D732R (SEQ ID NOS: 40-41), E39K/E742K (SEQ ID NOS: 42-43), and E39K/E189K (SEQ ID NOS: 44-45).
4. The mutant Taq polymerase of claim 3 wherein the remainder of the mutant polymerase has at least 95% amino acid sequence identity with wild type Taq polymerase.
5. The mutant Taq polymerase of claim 3 wherein the remainder of the mutant polymerase has at least 99% amino acid sequence identity with wild type Taq polymerase.
6. The mutant Taq polymerase of claim 3 wherein the remainder of the mutant polymerase has amino acid sequence identity with wild type Taq polymerase.
7. A mutant Taq polymerase comprising: one or more of the following amino acid mutations shown in the following even-numbered sequences, and wherein the remainder of the mutant polymerase has at least 80% amino acid sequence identity with wild type Taq polymerase, as shown in SEQ ID NO: 4 but wherein wild type Taq polymerase does not include the 6-membered histidine tag at its C-terminus and the six immediately preceding Glycine and Serine amino acids shown in SEQ ID NO: 4:
Q42R (SEQ ID NOS: 406-407), E101K (SEQ ID NOS: 46-47), V103S (SEQ ID NOS: 410-411), Y116A (SEQ ID NOS: 412-413), E117K (SEQ ID NOS: 48-49), E130K (SEQ ID NOS: 50-51), D144R (SEQ ID NOS: 6-7), E159K (SEQ ID NOS: 56-57), I163S (SEQ ID NOS: 404-405), E189C (SEQ ID NOS: 58-59), E189D (SEQ ID NOS: 60-61), E189K/E507K (SEQ ID NOS: 66-67), E189K/E537K (SEQ ID NOS: 68-69), E189K/D578R (SEQ ID NOS: 70-71), E189K/D732R (SEQ ID NOS: 72-73), and E189K/E742K (SEQ ID NOS: 74-75).
8. The mutant Taq polymerase of claim 7 wherein the remainder of the mutant polymerase has at least 95% amino acid sequence identity with wild type Taq polymerase.
9. The mutant Taq polymerase of claim 7 wherein the remainder of the mutant polymerase has at least 99% amino acid sequence identity with wild type Taq polymerase.
10. The mutant Taq polymerase of claim 7 wherein the remainder of the mutant polymerase has amino acid sequence identity with wild type Taq polymerase.