Mutant Taq polymerase for increased salt concentration or body fluids
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.
1 . 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:
V737S SEQ ID NOS: 438-439; E745K SEQ ID NOS: 466-467; M765S SEQ ID NOS: 468-469; E774K SEQ ID NOS: 470-471; L781S SEQ ID NOS: 472-473; A791F SEQ ID NOS: 474-475; E805K SEQ ID NOS: 476-477; L813S SEQ ID NOS: 478-479; Q534R SEQ ID NOS: 312-313; E537K SEQ ID NOS: 314-315; 1546S SEQ ID NOS: 356-357; and D547R SEQ ID NOS: 358-359.
2 . The mutant Taq polymerase of claim 1 wherein the PCR mixture further includes sodium.