Recombinant his-tagged DNA polymerase. II. Cloning and purification of Thermus aquaticus recombinant DNA polymerase (Stoffel fragment)

被引：7

作者：

Dabrowski, S ^{[1
]}

Kur, J ^{[1
]}

机构：

[1] Gdansk Univ Technol, Dept Microbiol, PL-80952 Gdansk, Poland

来源：

ACTA BIOCHIMICA POLONICA | 1998年 / 45卷 / 03期

关键词：

PCR; DNA polymerase; Thermus aquaticus; recombinant protein;

D O I：

10.18388/abp.1998_4204

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The Stoffel DNA fragment, shortened by 12 bp from 5' end, coding for Stoffel DNA polymerase (missing 4 amino acids at N-terminus of Stoffel amino-acids sequence) from the thermophilic Thermus aquaticus (strain YT-1) was amplified, cloned and expressed in Escherichia coli. The recombinant Stoffel fragment contained a polyhistidine tag at the N-terminus (21 additional amino acids) that allowed its single-step isolation by Ni2+ affinity chromatography. The enzyme was characterized and displayed high DNA polymerase activity and thermostability evidently higher than the native Tag DNA polymerase.

引用

页码：661 / 667

页数：7

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