兰州大学机构库 >第二临床医学院
大肠杆菌表达的TaqDNA聚合酶的纯化
Alternative TitlePurification of Taq DNA polymerase expressed in Escherichia coli
刘天磊; 薛守斌; 王芳; 朱琳颖; 梁微微; 曲圣轩; 蔡文博
2012-02-21
Source Publication遗传
ISSN0253-9772
Volume34Issue:3Pages:371-378
AbstractTaq DNA聚合酶是分子生物学研究中最常用的热稳定DNA聚合酶之一,与其他热稳定DNA聚合酶具有相似的特征,其纯化策略不但有潜在的应用前景,也对同类聚合酶的分离具有指导意义。已报道的适宜大量制备Taq酶的方案所需成本较高,而文章介绍了一种利用国产阳离子交换树脂廉价制备Taq酶的方案。在本方案中,采用热变性、(NH4)SO4沉淀与724离子交换层析分离大肠杆菌表达的Taq酶,约18 g Na型树脂干粉一次可回收比活约8 131.98 U/mg、总酶活2.2×105U、近27.07 mg Taq酶。纯化的产率可达48.92%,纯化倍数约59.35。所制酶SDS-PAGE电泳只检测到94 kDa单...
Other AbstractTaq 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.
KeywordTaqDNA聚合酶 724树脂 阳离子交换层析
Subject AreaBiochemistry & Molecular Biology ; Microbiology ; Genetics & Heredity
Publication PlaceBEIJING
Indexed ByPubMed ; MEDLINE ; BIOSIS ; CSCD
Language中文
First Inst
Host of Journal中国遗传学会 ; 中国科学院遗传与发育生物学研究所
Project Number江苏大学高级人才科研启动基金项目(编号:10JDG040),江苏大学百项本科生创新项目(编号:2010093)资助 ; 江苏高校优势学科建设工程资助项目(PAPD)
CSCD IDCSCD:4474469
PMID 22425957
BIOSIS IDBIOSIS:PREV201200349255
IRIDCNKI:0052214
Department江苏大学食品与生物工程学院;
兰州大学第二临床医学院
Citation statistics
Document Type期刊论文
Identifierhttp://ir.lzu.edu.cn/handle/262010/174313
Collection第二临床医学院
Recommended Citation
GB/T 7714
刘天磊,薛守斌,王芳,等. 大肠杆菌表达的TaqDNA聚合酶的纯化[J]. 遗传,2012,34(3):371-378.
APA 刘天磊.,薛守斌.,王芳.,朱琳颖.,梁微微.,...&蔡文博.(2012).大肠杆菌表达的TaqDNA聚合酶的纯化.遗传,34(3),371-378.
MLA 刘天磊,et al."大肠杆菌表达的TaqDNA聚合酶的纯化".遗传 34.3(2012):371-378.
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