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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Food Safety and Enteric Pathogens Research » Research » Publications at this Location » Publication #157693
Title: SLYA REGULATES THE COLLAGENASE-MEDIATED CYTOPATHIC PHENOTYPE IN MULTIRESISTANT SALMONELLA

Author
item Carlson, Steven
item McCuddin, Zoe
item WU, M

Submitted to: Microbial Pathogenesis
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/14/2005
Publication Date: 4/20/2005
Citation: Carlson, S.A., McCuddin, Z.P., Wu, M.T. 2005. SlyA regulates the collagenase-mediated cytopathic phenotype in multiresistant Salmonella. Microbial Pathogenesis. 38(4):181-187.

Interpretive Summary: Infections caused by multiple antibiotic resistant bacteria, especially Salmonella spp., in humans and livestock have become increasingly more prevalent. One of the major obstacles in studying these pathogens is that conventional genetic tools employing antibiotic selection are of little value since many of these isolates are resistant to most antibiotics. Here, we report a genetic system for strains that are multi-resistant. This protocol takes advantage of a transposon, a small motile piece of DNA, and the Zeocin**TM selection marker. Collectively, this protocol allows rapid, random disruptions to chromosomal genes and easy selection of mutants containing the insertions. Using this system we were able to assess genes in Salmonella that are involved in host invasion and antibiotic resistance. It will now be possible for scientists to identify novel genes involved in the hypervirulence of multiresistant Salmonella.

Technical Abstract: Infections caused by multiple antibiotic resistant bacteria, especially Salmonella spp., in humans and livestock have become increasingly more prevalent. One of the major obstacles in studying these pathogens is that conventional genetic tools employing antibiotic selection are of little value since many of these isolates are resistant to ampicillin, chloramphenicol, streptomycin, tetracycline, sulfamethoxazole, kanamycin, gentamicin, etc. Here, we report a modified mutagenesis protocol suitable for strains that are multi-resistant. This protocol takes advantage of the pMOD**TM transposon construction vector (Epicentre), the EZ::TN**TM transposon tool (Epicentre), and the Zeocin**TM selection marker (Invitrogen). Collectively, this protocol allows rapid, random disruptions to chromosomal genes and easy selection of mutants containing the insertions. With the EZ::TN mutagenesis followed by inverse PCR cloning, we were able to assess genes in Salmonella that are involved in host invasion and antibiotic resistance.