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Expression of recombinant proteins in the methane-producing archaeon Methanosarcina acetivorans

dc.contributor.advisorSowers, Kevin Ren_US
dc.contributor.authorMacAuley, Sheridan Roseen_US
dc.description.abstractRecombinant protein expression is a necessary tool for the investigation of proteins in the post-genomic era. While many systems exist for recombinant protein expression in organisms of the eukaryotic and eubacterial domains, few to none are available in the Archaea. A recombinant protein expression system using the methanogenic archaeon <em>Methanosarcina acetivorans</em> was developed which uses the highly regulated <em>cdh</em> promoter and allows expression of recombinant protein with optional 6xHis protein fusions to facilitate rapid purification. A protocol for high-density mass cultivation of <em>M. acetivorans</em> in a stainless steel bioreactor configured as a pH-auxostat was developed. The <em>cdh</em> promoter and alternate promoters were analyzed in attempt to enhance expression of recombinant proteins. The protein expression system was tested on several proteins: the <em>Methanocaldococcus jannaschii</em> prolyl tRNA synthetase, the <em>M. acetivorans</em> prolyl tRNA synthetase, the <em>Methanosarcina thermophila</em> carbonic anhydrase, and the <em>Dehalococcoides ethenogenes</em> tricholorethylene dehalogenase.en_US
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dc.titleExpression of recombinant proteins in the methane-producing archaeon Methanosarcina acetivoransen_US
dc.contributor.publisherDigital Repository at the University of Marylanden_US
dc.contributor.publisherUniversity of Maryland (College Park, Md.)en_US
dc.contributor.departmentMarine-Estuarine-Environmental Sciencesen_US
dc.subject.pqcontrolledBiology, Microbiologyen_US
dc.subject.pqcontrolledBiology, Geneticsen_US
dc.subject.pquncontrolledrecombinant proteinen_US
dc.subject.pquncontrolledoverexpression systemen_US

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