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dc.contributor.authorRussell, Christopher G.en
dc.date.accessioned2014-03-14T21:09:54Zen
dc.date.available2014-03-14T21:09:54Zen
dc.date.issued1993en
dc.identifier.otheretd-03022006-093402en
dc.identifier.urihttp://hdl.handle.net/10919/37446en
dc.description.abstractPast studies of mammary tissue specific expression of transgenes using the murine whey acidic protein (WAP) promoter have shown widely variable, positiondependent and copy number-dependent expression. This study evaluates a series of three WAP transgenes containing the cDNA of human protein C (hPC) for the expression of human protein C in the milk of mice. In two of the transgenes studied, the cDNA of (hPC) was inserted at the translational start site of a 7.8 kbp mouse WAP genomic DNA Eco RI fragment containing 2.6 kbp of 5’ flanking, 3.9 kbp WAP coding (exons and introns), and 1.3 kbp 3’ untranslated region (UTR) and flanking sequences (designated WAPPC1 and WAPPC2). A third transgene consisted of only the 2.6 kbp of WAP 5’ UTR and flanking DNA, 1.4 kbp hPC cDNA, and 1.3 kbp of 3’ WAP UTR and flanking DNA with no linker sequences (designated WAPPC3). The WAPPC1 and WAPPC2 transgenes expressed up to about 10 μg/ml recombinant hPC in mouse milk while WAPPC3 expressed 30-300 (n=10, n=5, n=11, number of founder lines evaluated for each transgene, respectively). In contrast to past studies with WAP-cDNA fusion transgenes where the maximal expression was about 5% of endogenous WAP expression, the WAPPC3 transgene gave maximal expression which was about 30% of endogenous WAP expression. Thus, results from the combination in WAPPC3 of intact 5’ and 3’ WAP UTR with the cDNA of hPC suggests that introns are not necessary to enable high level expression in the mammary gland when using WAP regulatory elements. Relative specific transcript and protein levels in the transgenic animals studied suggest that the rates of translation initiation may be different for the mRNAs of each of the transgenes studied.en
dc.format.extentviii, 149 leavesen
dc.format.mediumBTDen
dc.format.mimetypeapplication/pdfen
dc.language.isoenen
dc.publisherVirginia Techen
dc.relation.isformatofOCLC# 30698186en
dc.relation.haspartLD5655.V856_1993.R877.pdfen
dc.rightsIn Copyrighten
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en
dc.subject.lccLD5655.V856 1993.R877en
dc.subject.lcshGene expressionen
dc.subject.lcshMilk -- Genetic aspectsen
dc.subject.lcshProtein C -- Genetic aspectsen
dc.subject.lcshTransgenic miceen
dc.titleImprovement of expression of recombinant human protein C in the milk of transgenic animals using a novel transgene constructen
dc.typeDissertationen
dc.contributor.departmentChemical Engineeringen
dc.description.degreePh. D.en
thesis.degree.namePh. D.en
thesis.degree.leveldoctoralen
thesis.degree.grantorVirginia Polytechnic Institute and State Universityen
thesis.degree.disciplineChemical Engineeringen
dc.contributor.committeechairVelander, William H.en
dc.contributor.committeememberJohnson, John L.en
dc.contributor.committeememberWong, Eric A.en
dc.contributor.committeememberDavis, Richey M.en
dc.contributor.committeememberDrohan, William N.en
dc.type.dcmitypeTexten
dc.identifier.sourceurlhttp://scholar.lib.vt.edu/theses/available/etd-03022006-093402/en
dc.date.sdate2006-03-02en
dc.date.rdate2006-03-02en
dc.date.adate2006-03-02en


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