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dc.creatorSkretas, Georgiosen
dc.creatorGeorgiou, Georgeen
dc.date.accessioned2015-09-09T15:50:26Zen
dc.date.available2015-09-09T15:50:26Zen
dc.date.issued2010-09en
dc.identifier.citationSkretas, Georgios, Georgiou, George, >Simple Genetic Selection Protocol for Isolation of Overexpressed Genes That Enhance Accumulation of Membrane-Integrated Human G Protein-Coupled Receptors in Escherichia coli,> Appl. Environ. Microbiol. September 2010 vol. 76 no. 17 5852-5859. doi: 10.1128/AEM.00963-10.en
dc.identifier.issn0099-2240en
dc.identifier.urihttp://hdl.handle.net/2152/31084en
dc.description.abstractThe efficient production of membrane proteins in bacteria remains a major challenge. In this work, we sought to identify overexpressed genes that enhance the yields of recombinant membrane proteins in Escherichia coli. We developed a genetic selection system for bacterial membrane protein production, consisting of membrane protein fusions with the enzyme beta-lactamase and facile selection of high-production strains on ampicillin-containing media. This system was used to screen the ASKA library, an ordered library of plasmids encoding all the known E. coli open reading frames (ORFs), and several clones with the ability to accumulate enhanced amounts of recombinant membrane proteins were selected. Notably, coexpression of ybaB, a gene encoding a putative DNA-binding protein of unknown function, was found to enhance the accumulation of a variety of membrane-integrated human G protein-coupled receptors and other integral membrane proteins in E. coli by up to 10-fold. The results of this study highlight the power of genetic approaches for identifying factors that impact membrane protein biogenesis and for generating engineered microbial hosts for membrane protein production.en
dc.description.sponsorshipWelch Foundationen
dc.description.sponsorshipNational Institutes of Health NIH GM 55090en
dc.language.isoEnglishen
dc.rightsAdministrative deposit of works to Texas ScholarWorks: This works author(s) is or was a University faculty member, student or staff member; this article is already available through open access or the publisher allows a PDF version of the article to be freely posted online. The library makes the deposit as a matter of fair use (for scholarly, educational, and research purposes), and to preserve the work and further secure public access to the works of the University.en
dc.subjectgreen fluorescent proteinen
dc.subjectsignal recognition particleen
dc.subjectbeta-lactamaseen
dc.subjecthaemophilus-influenzaeen
dc.subjectexpressionen
dc.subjecttopologyen
dc.subjectbindingen
dc.subjectsystemen
dc.subjectexporten
dc.subjectpurificationen
dc.subjectbiotechnology & applied microbiologyen
dc.subjectmicrobiologyen
dc.titleSimple Genetic Selection Protocol For Isolation Of Overexpressed Genes That Enhance Accumulation Of Membrane-Integrated Human G Protein-Coupled Receptors In Escherichia Colien
dc.typeArticleen
dc.rights.holderen
dc.description.departmentCellular and Molecular Biologyen
dc.description.departmentChemical Engineeringen
dc.identifier.doi10.1128/aem.00963-10en
dc.identifier.urlen
dc.contributor.utaustinauthorSkretas, Georgiosen
dc.contributor.utaustinauthorGeorgiou, Georgeen
dc.relation.ispartofserialApplied and Environmental Microbiologyen


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