重组Taq DNA聚合酶在大肠杆菌中的表达与纯化

从嗜热菌Thermus aquaticus中分离出的Taq DNA聚合酶由于其热稳定性，在聚合酶链式反应(PCR)中得到广泛的应用，我们根据编码Taq DNA聚合酶的基因序列，设计出一对引物，通过PCR扩增出Taq DNA聚合酶基因，并将其克隆到表达载体pET-15b中，转化大肠杆菌E．coli BL21(DE3)，经IPTG诱导后，重组Taq DNA聚合酶在大肠杆菌中实现了大量表达。经过热变性、亲和层析、阳离子交换层析获得了电泳纯的重组Taq DNA聚合酶。本研究的生产程序可作为Taq DNA聚合酶生产的一种新方法。

Expression and Purification of Recombinant Taq DNA Polymerase in Escherichia Coli

Taq DNA polymerase from the thermophiles Thermus aquaticus is widely used in polymerase chain reaction because of its thermostability. The Taq Pol I gene from the genomic DNA of Thermus aquaticus YT-1 was amplified by PCR, then the amplified Taq Pol I gene was inserted into pET-15b to construct expressing vector pET-15bTaq. This expressing vector was introduced into E. coli BL21 (DE3), and the recombinant Taq DNA polymerase was expressed by IPTG induction. The recombinant protein was purified by heat treatment, Ni~(2 ) chelating resin affinity chromatography and chromatography on Sp-Sepharose column as well judged by SDS-PAGE.