Journal
PLASMID
Volume 59, Issue 3, Pages 231-237Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.plasmid.2008.01.001
Keywords
TEV protease-cleavable tags; rapid protein purification; propionate catabolism; 2-methylcitric acid cycle enzymes; 2-methylaconitate isomerase; 2-methylcitrate dehydratase
Categories
Funding
- NIAMS NIH HHS [AR35186, R01 AR035186-20, R01 AR035186, R01 AR035186-18, R01 AR035186-19, R01 AR035186-23, R01 AR035186-21, R01 AR035186-22] Funding Source: Medline
- NIGMS NIH HHS [R01 GM062203-07, GM62203, R01 GM062203] Funding Source: Medline
Ask authors/readers for more resources
We describe the construction and use of two sets of vectors for the over-expression and purification of protein from Escherichia coli. The set of pTEV plasmids (pTEV3, 4, 5) directs the synthesis of a recombinant protein with a N-terminal hexahistidine (HiS(6)) tag that is removable by the tobacco etch virus (TEV) protease. The set of pKLD plasmids (pKLD66, 116) directs the synthesis of a recombinant protein that contains a N-terminal HiS(6) and maltose-binding protein tag in tandem, which can also be removed with TEV protease. The usefulness of these plasmids is illustrated by the rapid, high-yield purification of the 2-methylcitrate dehydratase (PrpD) protein of Salmonella enterica, and the 2-methylaconitate isomerase (PrpF) protein of Shewanella oneidensis, two enzymes involved in the catabolism of propionate to pyruvate via the 2-methylcitric acid cycle. (c) 2008 Elsevier Inc. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available