【摘要】 Taq 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。试验利用含Taq DNA聚合酶基因的pTaq表达质粒转化大肠杆菌E.coli菌株,用异丙基硫代-β-D-乳糖苷(IPTG)诱导表达耐热Taq DNA聚合酶;采用热变性沉淀杂蛋白,SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)检测纯度,Bio-Rex 70柱层析纯化蛋白,PCR反应检测Taq DNA聚合酶的浓度和活性。结果表明:分离纯化制备的Taq DNA聚合酶,其纯度、酶活性和酶特异性均达到市售的Taq DNA聚合酶水平。更多还原

【Abstract】 Taq DNA polymerase is one of the most commonly used thermostable DNA polymerases in molecular biological research.The experiment used Taq DNA polymerase gene contained pTaq expression plasmid for transforming E.coli,and used IPTG for inducing expression of heat-tolerance Taq DNA polymerase.Moreover,it removed heteropolymeric protein by heat denaturation,examined purity by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE),and purified protein by Bio-Rex 70 column chromatography,and tested concentration and activity of the Taq DNA polymerase by polymerase chain reaction(PCR).The results revealed that the Taq DNA polymerase which was prepared and purified by this method could reach commercial level of Taq DNA polymerase in purity,enzyme activity and enzyme specificity.更多还原