Document Detail


A dual ELP-tagged split intein system for non-chromatographic recombinant protein purification.
MedLine Citation:
PMID:  23212673     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Self-cleaving elastin-like protein (ELP) tags provide a very promising tool for recombinant protein purification. With this method, the target protein is purified by simple ELP-mediated precipitation steps, followed by self-cleavage and removal of the ELP tag. Unfortunately, however, inteins usually experience some level of pre-cleavage during protein expression, which can significantly decrease final yields. In this study, we solve this problem by splitting the intein into two ELP-tagged segments. Each segment is incapable of pre-cleavage alone, but the assembled segments release the target protein rapidly when assembled in vitro. The result is the very tight control of the tag cleaving reaction, combined with the simplicity of the ELP purification method. Using this system, we successfully purified four different sizes of target proteins with final yields comparable to or higher than our original contiguous intein-ELP system. Further, we demonstrate a streamlined split intein method, where cells expressing the tagged intein segments are combined prior to cell lysis, allowing the segments to be co-purified in a single reaction mixture.
Authors:
Changhua Shi; Qing Meng; David W Wood
Related Documents :
10949663 - Cellular effects and signalling pathways activated by the anti-coagulant factor, protei...
23201283 - Overproduction of recombinant human mannose-binding lectin (mbl) in chinese hamster ova...
7692893 - Antigenic analysis of pseudomonas aeruginosa and pseudomonas cepacia groel proteins and...
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-12-5
Journal Detail:
Title:  Applied microbiology and biotechnology     Volume:  -     ISSN:  1432-0614     ISO Abbreviation:  Appl. Microbiol. Biotechnol.     Publication Date:  2012 Dec 
Date Detail:
Created Date:  2012-12-5     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8406612     Medline TA:  Appl Microbiol Biotechnol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Institute of Biological Sciences and Biotechnology, Donghua University, Shanghai, 201620, People's Republic of China.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Marine bacteria: potential candidates for enhanced bioremediation.
Next Document:  Expression of the immunoreactive buckwheat major allergenic storage protein in Lactococcus lactis.