Open AccessPurification of <I>Taq</I> DNA polymerase expressed in <I>Escherichia coli</I>
Author(s) -
Tian-Lei Liu,
Shoubin Xue,
Fang Wang,
Linying Zhu,
Weiwei Liang,
Sheng-Xuan Qu,
Wenbo Cai
Publication year - 2012
Publication title -
yichuan
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.125
H-Index - 16
ISSN - 0253-9772
DOI - 10.3724/sp.j.1005.2012.00371
Subject(s) - chromatography , microbiology and biotechnology , ammonium sulfate precipitation , chemistry , dna extraction , dna , taq polymerase , polymerase chain reaction , dna polymerase , polymerase , ammonium sulfate , polymerase chain reaction optimization , escherichia coli , enzyme , biology , biochemistry , gene , thermus aquaticus , nested polymerase chain reaction , size exclusion chromatography
Taq DNA polymerase is one of the most commonly thermostable DNA polymerases in molecular biological researches, which shares its basic characters with others of the family, thereby its purifying strategy could be used not only in itself production but also in the extraction of the others as a reference. At present, the protocols reported for large scale preparation of Taq DNA are high cost, so a cheaper method was described here. In this protocol, by heat denaturation, ammonium sulfate precipitation and cation exchange chromatography of 724 resin, about 18 g powder of Na form resin could recover about 27.07 mg of Taq enzyme. The total activity and specific activity were approximately 2.2 × 10⁵ U and 8131.98 U/mg. The total yield was about 48.92% with 59.35 of purification folds. Analysis of quality of purified enzyme indicated that only one protein 94 kDa was identified against SDS-PAGE and the remnant of DNA nuclease was not detected. For PCR reaction, The amplification ability of purified Taq polymerase was not different from that of the commercially avail-able ones. This method reported in the present study is effective and low cost, making it suitable for general purification in laboratories or business production.