Linked Life Data

17948006

Source:http://linkedlifedata.com/resource/pubmed/id/17948006

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
10
pubmed:dateCreated
2007-10-19
pubmed:abstractText
Eight combinations of molecular chaperones (e.g., DnaK/DnaJ/GrpE/ClpB) are co-expressed with the target recombinant protein to compare their effectiveness in improving its soluble yield. This system allows the most complete and rational approach proposed so far to use the chaperone activity for optimizing the host cell folding machinery. Furthermore, a two-step protocol is presented, in which protein synthesis and protein refolding are uncoupled. Molecular chaperones and target protein accumulate in the first growth phase and target protein aggregates are then disaggregated in vivo after the block of protein synthesis. The optimal chaperone combination to maximize the soluble yield of a specific protein remains unpredictable. Therefore, a small-scale purification selection step is useful for screening among expression combinations before scaling-up production. Applying such a strategy, we could increase the solubility of 70% of the tested constructs with yields of up to 42-fold more protein than in controls. The procedure takes 2 d.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:issn
1750-2799
pubmed:author
pubmed:issnType
Electronic
pubmed:volume
2
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
2632-9
pubmed:dateRevised
2008-3-24
pubmed:meshHeading
pubmed:year
2007
pubmed:articleTitle
Protocol for preparing proteins with improved solubility by co-expressing with molecular chaperones in Escherichia coli.
pubmed:affiliation
Scientific Core Facilities, EMBL, Meyerhofstrasse 1, D-69117 Heidelberg, Germany. ario.demarco@ifom-ieo-campus.it
pubmed:publicationType
Journal Article