Toggle Main Menu Toggle Search

Open Access padlockePrints

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

Lookup NU author(s): Isa Gokce, Professor Jeremy LakeyORCiD

Downloads

Full text for this publication is not currently held within this repository. Alternative links are provided below where available.


Abstract

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 His6-Ser2 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. © 2002 Elsevier Science (USA). All rights reserved.


Publication metadata

Author(s): Anderluh G, Gokce I, Lakey JH

Publication type: Article

Publication status: Published

Journal: Protein Expression and Purification

Year: 2003

Volume: 28

Issue: 1

Pages: 173-181

ISSN (print): 1046-5928

ISSN (electronic): 1096-0279

Publisher: Academic Press

URL: http://dx.doi.org/10.1016/S1046-5928(02)00681-2

DOI: 10.1016/S1046-5928(02)00681-2

PubMed id: 12651122


Altmetrics

Altmetrics provided by Altmetric


Share