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dc.contributor.authorMatic, Jake N
dc.contributor.authorTerry, Tamsin D
dc.contributor.authorVan Bockel, David
dc.contributor.authorMaddocks, Tracy
dc.contributor.authorTinworth, David
dc.contributor.authorJennings, Michael P
dc.contributor.authorDjordjevic, Steven P
dc.contributor.authorWalker, Mark J
dc.date.accessioned2017-05-03T15:42:28Z
dc.date.available2017-05-03T15:42:28Z
dc.date.issued2009
dc.date.modified2010-09-10T05:18:14Z
dc.identifier.issn0019-9567
dc.identifier.doi10.1128/IAI.01301-08
dc.identifier.urihttp://hdl.handle.net/10072/33885
dc.description.abstractLive-vaccine delivery systems expressing two model antigens from Mycoplasma hyopneumoniae, F2P97 (Adh) and NrdF, were constructed using Salmonella enterica serovar Typhimurium aroA (STM-1), and immunogenicity in mice was evaluated. Recombinant plasmid-based expression (PBE) and chromosomally based expression (CBE) systems were constructed. The PBE system was formed by cloning both antigen genes into pJLA507 to create an operon downstream of temperature-inducible promoters. Constitutive CBE was achieved using a promoter-trapping technique whereby the promoterless operon was stably integrated into the chromosome of STM-1, and the expression of antigens was assessed. The chromosomal position of the operon was mapped in four clones. Inducible CBE was obtained by using the in vivo-induced sspA promoter and recombining the expression construct into aroD. Dual expression of the antigens was detected in all systems, with PBE producing much larger quantities of both antigens. The stability of antigen expression after in vivo passage was 100% for all CBE strains recovered. PBE and CBE strains were selected for comparison in a vaccination trial. The vaccine strains were delivered orally into mice, and significant systemic immunoglobulin M (IgM) and IgG responses against both antigens were detected among all CBE groups. No significant immune response was detected using PBE strains. Expression of recombinant antigens in S. enterica serovar Typhimurium aroA from chromosomally located strong promoters without the use of antibiotic resistance markers is a reliable and effective method of inducing a significant immune response.
dc.description.peerreviewedYes
dc.description.publicationstatusYes
dc.languageEnglish
dc.language.isoeng
dc.publisherAmerican Society for Microbiology
dc.publisher.placeUnited States
dc.relation.ispartofstudentpublicationN
dc.relation.ispartofpagefrom1817
dc.relation.ispartofpageto1826
dc.relation.ispartofissue5
dc.relation.ispartofjournalInfection and Immunity
dc.relation.ispartofvolume77
dc.rights.retentionY
dc.subject.fieldofresearchBiological sciences
dc.subject.fieldofresearchInfectious agents
dc.subject.fieldofresearchAgricultural, veterinary and food sciences
dc.subject.fieldofresearchBiomedical and clinical sciences
dc.subject.fieldofresearchcode31
dc.subject.fieldofresearchcode310702
dc.subject.fieldofresearchcode30
dc.subject.fieldofresearchcode32
dc.titleDevelopment of Non-Antibiotic-Resistant, Chromosomally Based, Constitutive and Inducible Expression Systems for aroA-Attenuated Salmonella enterica Serovar Typhimurium
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
gro.date.issued2009
gro.hasfulltextNo Full Text
gro.griffith.authorJennings, Michael P.


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