Mutant Taq Polymerase for Increased Salt Concentration Use
AssigneeAbClonal Science, Inc.
InventorsZhenyu Zhu, Dapeng Sun, Weiling Peng
Life of this patent
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Abstract
The invention includes a mutant Taq polymerase, which can effectively amplify a target sequence under conditions of salt concentration(s) similar to body fluids, including blood, serum or plasma preserved with sodium citrate. 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.
Claims20
A mutant Taq polymerase which is active in a whole blood, plasma or serum, or in a PCR mixture which includes KCl at a concentration of 10 to 110 mM, comprising one of the following amino acid mutations preceding an even-numbered sequence identification number but wherein each said amino acid sequence does not include the 6-membered histidine tag at its C-terminus and the six immediately preceding Glycine and Serine amino acids shown in each said amino acid sequence, and wherein a DNA encoding each said amino acid sequence is the odd-numbered sequence identification number following each said even-numbered sequence identification number:
- I553S SEQ ID NOS:
- 366-367;
- P555G SEQ ID NOS:
- 368-369;
- N565R SEQ ID NOS:
- 370-371;
- D578R SEQ ID NOS:
- 372-373;
- G603P SEQ ID NOS:
- 400-401;
- E681K SEQ ID NOS:
- 402-403;
- L682S SEQ ID NOS:
- 404-405;
- P701G SEQ ID NOS:
- 408-409;
- K702E SEQ ID NOS:
- 410-411;
- L233S SEQ ID NOS:
- 178-179;
- V256S SEQ ID NOS:
- 180-181;
- E288K SEQ ID NOS:
- 186-187;
- E303K SEQ ID NOS:
- 188-189;
- P336G SEQ ID NOS:
- 200-201;
- E337K SEQ ID NOS:
- 202-203;
- L351S SEQ ID NOS:
- 208-209;
- P373G SEQ ID NOS:
- 216-217;
- D381R SEQ ID NOS:
- 218-219;
- and D452R SEQ ID NOS:
- 222-223.
A mutant Taq polymerase which is active in a whole blood, plasma or serum, or in a PCR mixture which includes KCl at a concentration of 10 to 110 mM, comprising one of the following amino acid mutations preceding a comma, at the positions indicated in each amino acid sequence and as fully shown in an adjacent even-numbered sequence identification number preceding a comma but wherein each said amino acid sequence does not include the 6-membered histidine tag at its C-terminus and the six immediately preceding Glycine and Serine amino acids shown in each said amino acid sequence, and wherein a DNA encoding each said amino acid sequence is the odd-numbered sequence identification number following each said even-numbered sequence identification number preceding a comma:
- E520K SEQ ID NOS:
- 268-269;
- A521F SEQ ID NOS:
- 310-311;
- D91R SEQ ID NOS:
- 28-29;
- D104R SEQ ID NOS:
- 32-33;
- L111S SEQ ID NOS:
- 34-35;
- T186I SEQ ID NOS:
- 36-37;
- E201K SEQ ID NOS:
- 112-113;
- E215K SEQ ID NOS:
- 116-117;
- K219E SEQ ID NOS:
- 118-119;
- L221S SEQ ID NOS:
- 120-121;
- D222R SEQ ID NOS:
- 122-123;
- L365S SEQ ID NOS:
- 210-211;
- G366P SEQ ID NOS:
- 212-213;
- P368G SEQ ID NOS:
- 214-215;
- N384R SEQ ID NOS:
- 220-221;
- L541S SEQ ID NOS:
- 354-355;
- P550G SEQ ID NOS:
- 360-361;
- and L552S SEQ ID NOS:
- 364-365.
Dependencies are inferred from the claim text.