Linked Life Data

20542040

Source:http://linkedlifedata.com/resource/pubmed/id/20542040

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2010-7-19
pubmed:abstractText
The production of complex multidomain (membrane) proteins is a major hurdle in structural genomics and a generic approach for optimizing membrane protein expression is still lacking. We have devised a selection method to isolate mutant strains with improved functional expression of recombinant membrane proteins. By fusing green fluorescent protein and an erythromycin resistance marker (ErmC) to the C-terminus of a target protein, one simultaneously selects for variants with enhanced expression (increased erythromycin resistance) and correct folding (green fluorescent protein fluorescence). Three evolved hosts, displaying 2- to 8-fold increased expression of a plethora of proteins, were fully sequenced and shown to carry single-site mutations in the nisK gene. NisK is the sensor protein of a two-component regulatory system that directs nisin-A-mediated expression. The levels of recombinant membrane proteins were increased in the evolved strains, and in some cases their folding states were improved. The generality and simplicity of our approach allow rapid improvements of protein production yields by directed evolution in a high-throughput way.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
1089-8638
pubmed:author
pubmed:copyrightInfo
Copyright (c) 2010 Elsevier Ltd. All rights reserved.
pubmed:issnType
Electronic
pubmed:day
6
pubmed:volume
401
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
45-55
pubmed:meshHeading
pubmed-meshheading:20542040-Cloning, Molecular, pubmed-meshheading:20542040-Directed Molecular Evolution, pubmed-meshheading:20542040-Gene Expression Regulation, Bacterial, pubmed-meshheading:20542040-Genes, Bacterial, pubmed-meshheading:20542040-Genes, Reporter, pubmed-meshheading:20542040-Genome, Bacterial, pubmed-meshheading:20542040-Green Fluorescent Proteins, pubmed-meshheading:20542040-High-Throughput Screening Assays, pubmed-meshheading:20542040-Lactococcus lactis, pubmed-meshheading:20542040-Membrane Proteins, pubmed-meshheading:20542040-Organisms, Genetically Modified, pubmed-meshheading:20542040-Protein Folding, pubmed-meshheading:20542040-Recombinant Fusion Proteins, pubmed-meshheading:20542040-Recombinant Proteins, pubmed-meshheading:20542040-Species Specificity, pubmed-meshheading:20542040-Up-Regulation
pubmed:year
2010
pubmed:articleTitle
Evolved Lactococcus lactis strains for enhanced expression of recombinant membrane proteins.
pubmed:affiliation
Department of Biochemistry, Groningen Biomolecular Sciences and Biotechnology Institute, Netherlands Proteomics Centre, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't