Author/Editor		Anderluh, Gregor; Goekce, Isa; Lakey, Jeremy H
Title		Expression of proteins using the third domain of the Escherichia coli periplasmic-protein TolA as a fusion partner
Type		članek
Source		Protein Expr Purif
Vol. and No.		Letnik 28
Publication year		2003
Volume		str. 173-81
Language		eng
Abstract		The third domain of the periplasmic protein To1A from Escherichia coli (To1AIII) was used as a fusion partner in the expression of various proteins from bacteria and eukaryotes. To1AIII 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 To1AIII 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 His6Ser2 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.
Descriptors		ESCHERICHIA COLI CARRIER PROTEINS CLONING, MOLECULAR BACTERIAL PROTEINS COLICINS CNIDARIAN VENOMS THROMBIN FACTOR XA BASE SEQUENCE