title: Activity of native vs. synthetic promoters in Brucella.
abstract: Brucellosis caused by Brucella species is reportedly the most common zoonotic infection worldwide. The bacterial pathogen is also classified by the Centers for Disease Control and Prevention as a category (B) pathogen that has the potential for development as a bioweapon. Although eight genomes of Brucella have been sequenced, little information is available regarding the regulation of gene expression and promoter activity in Brucella spp. We therefore constructed a set of broad-host-range vectors expressing the lacZ reporter gene from various promoters. Four groups of promoters (Brucella native, antibiotic resistant, bacteriophage and synthetic promoters) were tested in vivo and in vitro in Brucella suis. The highest level of heterologous gene expression was achieved with synthetic hybrid trc promoter carrying the adenine-rich upstream element. Furthermore, this demonstrates the usefulness of synthetic promoters for enhanced level of gene expression in Brucella spp.
authors.affiliation: Institute for Critical Technology and Applied Science, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA.
authors.forename: Mohamed N
authors.initials: MN
authors.lastname: Seleem
authors.forename: Neeta
authors.initials: N
authors.lastname: Jain
authors.forename: Hamzeh
authors.initials: H
authors.lastname: Alqublan
authors.forename: Ramesh
authors.initials: R
authors.lastname: Vemulapalli
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Boyle
authors.forename: Nammalwar
authors.initials: N
authors.lastname: Sriranganathan
mesh_terms: D000818:Animals; D041021:Brucella suis / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D005822:Genetic Vectors; D007763:Lac Operon; D008264:Macrophages / Q000382:microbiology; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics* / Q000502:physiology*; D017422:Sequence Analysis, DNA; D001616:beta-Galactosidase / Q000235:genetics* / Q000378:metabolism*
chemical_list: D001616:beta-Galactosidase