Expression of proteins using the third domain of the Escherichia coli periplasmic-protein TolA as a fusion partner

被引:31
作者
Anderluh, G
Gökçe, I
Lakey, JH
机构
[1] Univ Newcastle Upon Tyne, Sch Cellular & Mol Biosci, Newcastle Upon Tyne NE2 4HH, Tyne & Wear, England
[2] Univ Ljubljana, Dept Biol, Ljubljana 1000, Slovenia
[3] Gaziosmanpasa Univ, Dept Chem, Tokat, Turkey
基金
英国生物技术与生命科学研究理事会; 英国惠康基金;
关键词
D O I
10.1016/S1046-5928(02)00681-2
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The third domain of the periplasmic protein TolA from Escherichia coli (TolAIII) was used as a fusion partner in the expression of various proteins from bacteria and eukaryotes. TolAIII is small domain, expressed in high yields as a soluble protein in the cytoplasm of E coli. Proteins were linked to the C-terminus of TolAIII by a short flexible linker containing sites for endopeptidases. Three different vectors were prepared, containing sites for enterokinase, thrombin or factor Xa. Fusion proteins also contain a His(6)-Ser(2) tag at their N-terminus for easier purification. Up to 90 mg fusion protein per liter bacterial culture was obtained using these vectors. Colicin N R-domain was expressed with this system as a fusion and processed further for functional studies. The yield of final pure R-domain was doubled as compared to the direct expression. The system may prove to be useful in the preparation of other peptides and proteins. (C) 2002 Elsevier Science (USA). All rights reserved.
引用
收藏
页码:173 / 181
页数:9
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