Tuning Gene Expression In Yarrowia Lipolytica By A Hybrid Promoter Approach

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dc.contributor.utaustinauthorBlazeck, Johnen
dc.contributor.utaustinauthorLiu, Leqianen
dc.contributor.utaustinauthorRedden, Heidien
dc.contributor.utaustinauthorAlper, Halen
dc.creatorBlazeck, Johnen
dc.creatorLiu, Leqianen
dc.creatorRedden, Heidien
dc.creatorAlper, Halen
dc.date.accessioned2015-09-09T15:50:20Zen
dc.date.available2015-09-09T15:50:20Zen
dc.date.issued2011-11en
dc.description.abstractThe development of strong and tunable promoter elements is necessary to enable metabolic and pathway engineering applications for any host organism. Here, we have expanded and generalized a hybrid promoter approach to produce libraries of high-expressing, tunable promoters in the nonconventional yeast Yarrowia lipolytica. These synthetic promoters are comprised of two modular components: the enhancer element and the core promoter element. By exploiting this basic promoter architecture, we have overcome native expression limitations and provided a strategy for both increasing the native promoter capacity and producing libraries for tunable gene expression in a cellular system with ill-defined genetic tools. In doing so, this work has created the strongest promoters ever reported for Y. lipolytica. Furthermore, we have characterized these promoters at the single-cell level through the use of a developed fluorescence-based assay as well as at the transcriptional and whole-cell levels. The resulting promoter libraries exhibited a range of more than 400-fold in terms of mRNA levels, and the strongest promoters in this set had 8-fold-higher fluorescence levels than those of typically used endogenous promoters. These results suggest that promoters in Y. lipolytica are enhancer limited and that this limitation can be partially or fully alleviated through the addition of tandem copies of upstream activation sequences (UASs). Finally, this work illustrates that tandem copies of UAS regions can serve as synthetic transcriptional amplifiers that may be generically used to increase the expression levels of promoters.en
dc.description.departmentCellular and Molecular Biologyen
dc.description.departmentChemical Engineeringen
dc.description.sponsorshipDuPonten
dc.identifier.citationBlazeck, John, Liu, Lequin, Redden, Heidi, Alper, Hal, >Tuning gene expression in Yarrowia lipolytica by a hybrid promoter approach,> Appl Environ Microbiol. 2011 Nov;77(22):7905-14. doi: 10.1128/AEM.05763-11.en
dc.identifier.doi10.1128/aem.05763-11en
dc.identifier.issn0099-2240en
dc.identifier.urihttp://hdl.handle.net/2152/31074en
dc.identifier.urlen
dc.language.isoEnglishen
dc.relation.ispartofserialApplied and Environmental Microbiologyen
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.rights.holderen
dc.subjectsaccharomyces-cerevisiaeen
dc.subjectheterologous proteinsen
dc.subjectlipid-accumulationen
dc.subjectxpr2 geneen
dc.subjectyeasten
dc.subjectsequencesen
dc.subjectcassettesen
dc.subjectvectorsen
dc.subjectregionsen
dc.subjectcloningen
dc.subjectbiotechnology & applied microbiologyen
dc.subjectmicrobiologyen
dc.titleTuning Gene Expression In Yarrowia Lipolytica By A Hybrid Promoter Approachen
dc.typeArticleen

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