Expression and characterization of recombinant bovine C3d fusion proteins

dc.contributor.advisorShewen, Patricia E.
dc.contributor.authorFirth, Matthew A. of Pathobiologyen_US of Guelphen_US of Scienceen_US
dc.description.abstractThe gene fragment encoding bovine complement component C3d (boC3d) was isolated, cloned, sequenced and expressed in preparation for investigation of its potential as an adjuvant in cattle. Studies in mice attribute the adjuvant activity of C3d to a lowered B-cell activation threshold, resulting from the cross-linking of membrane-Ig and the B-cell co-receptor complex CD21(CR2)/CD19/CD81. To evaluate binding of recombinant boC3d to native bovine CD21, a recombinant boC3d protein was generated that included a biotinylation signal sequence at the N-terminal end of the boC3d sequence, for endogenous biotinylation by 'E. coli' via the BirA holoenzyme synthetase. Three additional constructs containing a portion of the gene encoding the leukotoxin produced by 'Mannheimia haemolytica' A1 linked to one, two or three boC3d units were created for expression in 'E. coli' as recombinant fusion proteins, as potential vaccines. All recombinant proteins incorporated polyhistidine tags and were purified by Ni-NTA agarose chromatography, then identified by SDS-PAGE and western blot. These proteins will provide the basis for future studies of receptor binding and cell-activation, and for immunization trials in cattle.en_US
dc.publisherUniversity of Guelphen_US
dc.rights.licenseAll items in the Atrium are protected by copyright with all rights reserved unless otherwise indicated.
dc.subjectrecombinant boC3den_US
dc.subjectbovine CD21en_US
dc.subjectboC3d proteinen_US
dc.subjectbiotinylation signal sequenceen_US
dc.subjectrecombinant fusion proteinsen_US
dc.subjectreceptor bindingen_US
dc.titleExpression and characterization of recombinant bovine C3d fusion proteinsen_US


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