Development of non-antibiotic-resistant, chromosomally based, constitutive and inducible expression systems for aroA-attenuated Salmonella enterica serovar typhimurium

Matic, Jake N., Terry, Tamsin D., Van Bockel, David, Maddocks, Tracy, Tinworth, David, Jennings, Michael P., Djordjevic, Steven P. and Walker, Mark J. (2009) Development of non-antibiotic-resistant, chromosomally based, constitutive and inducible expression systems for aroA-attenuated Salmonella enterica serovar typhimurium. Infection and Immunity, 77 5: 1817-1826. doi:10.1128/IAI.01301-08

Attached Files (Some files may be inaccessible until you login with your UQ eSpace credentials)
Name Description MIMEType Size Downloads
UQ181201_OA.pdf Full text (open access) application/pdf 691.14KB 0

Author Matic, Jake N.
Terry, Tamsin D.
Van Bockel, David
Maddocks, Tracy
Tinworth, David
Jennings, Michael P.
Djordjevic, Steven P.
Walker, Mark J.
Title Development of non-antibiotic-resistant, chromosomally based, constitutive and inducible expression systems for aroA-attenuated Salmonella enterica serovar typhimurium
Formatted title
Development of non-antibiotic-resistant, chromosomally based, constitutive and inducible expression systems for aroA-attenuated Salmonella enterica serovar typhimurium
Journal name Infection and Immunity   Check publisher's open access policy
ISSN 1098-5522
1070-6313
Publication date 2009-05
Sub-type Article (original research)
DOI 10.1128/IAI.01301-08
Open Access Status File (Publisher version)
Volume 77
Issue 5
Start page 1817
End page 1826
Total pages 10
Place of publication Washington, DC, United States
Publisher American Society for Microbiology
Collection year 2010
Language eng
Formatted abstract
Live-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.
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
Keyword Hyopneumoniae cilium adhesin
Mycoplasma-hyopneumoniae
Escherichia-coli
Vaccine strains
Var.-typhimurium
Bordetella-bronchiseptica
Heterologous antigens
Glycogen-synthesis
Stable expression
Oral immunization
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: 2010 Higher Education Research Data Collection
School of Chemistry and Molecular Biosciences
 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 13 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 19 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Thu, 03 Sep 2009, 08:18:33 EST by Mr Andrew Martlew on behalf of School of Chemistry & Molecular Biosciences