USING ANTISENSE MESSENGER RNA TO DOWNREGULATE LON MEDIATED PROTEOLYSIS IN ESCHERICHIA COLI

dc.contributor.advisorBentley, William Een_US
dc.contributor.authorCarter, Karenen_US
dc.contributor.departmentChemical Engineeringen_US
dc.date.accessioned2004-05-31T19:49:25Z
dc.date.available2004-05-31T19:49:25Z
dc.date.issued2003-12-18en_US
dc.description.abstractThe advent of metabolic engineering has instigated the introduction of foreign heterologous proteins into host cells, such as Escherichia coli. However, the metabolic burden incurred by the host cell to produce the desired recombinant protein elicits a cellular stress response that can result in reduced yields and degradation of the desired protein. In, lon is one of the major proteases responsible for this abnormal protein degradation, including recombinant proteins. Consequently, a variety of antisense strategies have been examined and shown to effectively control endogenous gene expression and function in E. coli. For this investigation we explored using a 300 base pair sequence of the 5' coding region of E. coli lon gene, including the start codon, cloned into both the pSE420 and pTO plasmids in the antisense (reverse) orientation. We examined the ability of lon antisense RNA to inhibit the production of endogenous lon protease and increase the protein yield and activity of a model recombinant protein, organophosphorus hydrolase (OPH). Results indicate that the overproduction of lon antisense did effectively downregulate the production of endogenous lon. In addition, cultures induced for lon antisense also revealed higher OPH protein levels in the first hour of production and a 7-fold higher activity.en_US
dc.format.extent1026222 bytes
dc.format.mimetypeapplication/pdf
dc.identifier.urihttp://hdl.handle.net/1903/108
dc.language.isoen_US
dc.relation.isAvailableAtDigital Repository at the University of Marylanden_US
dc.relation.isAvailableAtUniversity of Maryland (College Park, Md.)en_US
dc.subject.pqcontrolledEngineering, Chemicalen_US
dc.titleUSING ANTISENSE MESSENGER RNA TO DOWNREGULATE LON MEDIATED PROTEOLYSIS IN ESCHERICHIA COLIen_US
dc.typeThesisen_US

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