Purification and characterization of the 5′ → 3′ exonuclease domain-deleted Thermus filiformis DNA polymerase expressed in Escherichia coli
作者： Jeong Jin Choi , Hyun-Kyu Kim , Suk-Tae Kwon
关键词: Exonuclease 、 DNA polymerase II 、 Biochemistry 、 Recombinant DNA 、 Molecular biology 、 DNA polymerase 、 Escherichia coli 、 Molecular cloning 、 Klenow fragment 、 Thermus 、 Biology
摘要: The gene encoding 5′ → 3′ exonuclease domain-deleted Tfi DNA polymerase, named Exo−Tfi fragment, from Thermus filiformis was expressed in Escherichia coli under the control of tac promoter on a high-copy plasmid, pJR. enzyme purified 27-fold with 19% yield and specific activity 2621 U mg−1 protein. domain polymerase removed without significant effect stability. PCR conditions for fragment were more tolerant to buffer composition as compared full-length enzyme.
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