Journal: mSphere

Article Title: Role of RpoS in Regulating Stationary Phase Salmonella Typhimurium Pathogenesis-Related Stress Responses under Physiological Low Fluid Shear Force Conditions

doi: 10.1128/msphere.00210-22

Figure Lengend Snippet: Construction of rpoS mutant and molecular and phenotypic strain validation. (A) Deletion of the rpoS gene from the S. Typhimurium χ3339 genome; deletion start site +1, end site +993 bp. In the absence of an available sequenced genome for χ3339, the sequenced genome of strain SL1344 (the parental strain of χ3339) was used as a proxy. (B) Suicide vector pYA4804 was engineered by cloning a 717 bp fragment encompassing up-and downstream regions of the rpoS gene from S. Typhimurium χ3761 (wild-type UK-1) into the XbaI-SacI cloning sites of pRE112. (C) PCR analysis was used to confirm the deletion of rpoS . Lane 1, wild-type strain χ3339; lane 2, PCR negative control; lane 3, NEB 100 bp DNA ladder; lanes 4 and 5 show strains that did not have a successful double-crossover event to delete the rpoS gene and, thus, were not selected for use in this study. Lane 6 shows no PCR amplification of rpoS , thus demonstrating the successful deletion of the rpoS gene by the 2nd crossover. This strain was selected as the rpoS mutant for use in this study. (D) The χ3339 rpoS mutant exhibited reduced catalase activity compared to the wild-type strain.

Article Snippet: To construct this mutant, we used recombinant suicide vector pYA4804 (5.89 kb) carrying DNA fragments from S. Typhimurium strain χ3761 (wild-type UK-1) up-and downstream regions of the rpoS gene inserted into the XbaI-SacI cloning sites of vector pRE112 (Addgene) ( ).

Techniques: Mutagenesis, Plasmid Preparation, Clone Assay, Negative Control, Amplification, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Atlantic Salmon Mucins Inhibit LuxS -Dependent A. Salmonicida AI-2 Quorum Sensing in an N -Acetylneuraminic Acid-Dependent Manner

doi: 10.3390/ijms23084326

Figure Lengend Snippet: Strains and plasmids in this study.

Article Snippet: pRE112 plasmid , Suicide vector; R6K ori sacB Cm R , Addgene.

Techniques: Plasmid Preparation, Conjugation Assay