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
收藏
页码：661 / 667
页数：7
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