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dc.contributor.advisorMantle, Tim
dc.contributor.authorDunne, Aisling
dc.date.accessioned2018-12-06T11:56:55Z
dc.date.available2018-12-06T11:56:55Z
dc.date.issued2001
dc.identifier.citationAisling Dunne, 'Studies on human biliverdin-IX alpha reductase and linear tetrapyrrole signaling', [thesis], Trinity College (Dublin, Ireland). School of Biochemistry and Immunology, 2001, pp 259
dc.identifier.otherTHESIS 6353
dc.identifier.urihttp://hdl.handle.net/2262/85466
dc.description.abstractHuman Biliverdin-IXa reductase (BVR-A) has been cloned and overexpressed in E.coli as a GST- and Hexahistidine fusion protein. The full length cDNA encoding the enzyme has been amplified via PCR and hgated into the pGEX-KG and pTrcHis B expression vectors in order to produce the respective fusions. Induction of TGI cells transformed with the pGEX-BVR-A and pTrc-BVR-A constructs has culminated in the expression of a recombinant protein of the correct size and antigenicity in both cases. Purification of the GST-fusion protein on a glutathione affinity resin yields approximately 40 mg of fusion protein per litre of culture. The fusion protein has a molecular weight of 66 kDa, however, it is possible to remove the GST tag using the proteolytic enzyme, thrombin. The purified hBVR-A protein migrates on SDS-PAGE with a mobility corresponding to 40 kDa, however, mass spectroscopic analysis has confirmed the true relative molecular mass to be 34 kDa. A selenomethionine derivative of the recombinant hBVR-A protein has been prepared for use in Multiwavelength Anomalous Diffraction experiments and crystals diffracting at 3A have recently been obtained.
dc.format1 volume
dc.language.isoen
dc.publisherTrinity College (Dublin, Ireland). School of Biochemistry and Immunology
dc.relation.isversionofhttp://stella.catalogue.tcd.ie/iii/encore/record/C__Rb12462290
dc.subjectBiochemistry, Ph.D.
dc.subjectPh.D. Trinity College Dublin
dc.titleStudies on human biliverdin-IX alpha reductase and linear tetrapyrrole signaling
dc.typethesis
dc.type.supercollectionthesis_dissertations
dc.type.supercollectionrefereed_publications
dc.type.qualificationlevelDoctoral
dc.type.qualificationnameDoctor of Philosophy (Ph.D.)
dc.rights.ecaccessrightsopenAccess
dc.format.extentpaginationpp 259
dc.description.noteTARA (Trinity’s Access to Research Archive) has a robust takedown policy. Please contact us if you have any concerns: rssadmin@tcd.ie
dc.description.notePrint thesis water damaged as a result of the Berkeley Library Podium flood 25/10/2011


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