pbad24 vector  (ATCC)

Journal: Infection and Immunity

Article Title: A Salmonella enterica serovar Typhimurium genome-wide CRISPRi screen reveals a role for type 1 fimbriae in evasion of antibody-mediated agglutination

doi: 10.1128/iai.00574-24

Figure Lengend Snippet: Overexpression of T1F reduces STm susceptibility to Sal4-mediated agglutination in the snow globe assay. ( A ) Recovered CFU/mL of STm WT + pBAD24-EV (empty vector; EV), Δ fimW + EV, and Δ fimW + pBAD24- fimW (pFimW) cultures in the snow globe assay. ( B ) Quantification of mannose-sensitive yeast agglutination of the STm WT + EV, Δ fimW + EV, and Δ fimW + pFimW strains. ( C ) Recovered CFU/mL of STm WT + EV, Δ fimW + EV, Δ fimA + EV, and Δ fimA + pFimW cultures in the snow globe assay. ( D ) Quantification of mannose-sensitive yeast agglutination of WT + EV, Δ fimW + EV, Δ fimA + EV, and Δ fimA + pFimW strains. For panels A and C, the indicated strains were grown to mid-log phase in the presence of 0.02% arabinose, washed in PBS, and either left untreated (circles) or treated with 15 μg/mL of Sal4 IgA (squares). After 2 h of treatment, the top of the supernatant was collected and plated on LB agar to measure CFU. For panels B and D, the indicated strains were incubated statically for 48 h in LB containing 0.02% arabinose at 37°C prior to centrifugation and resuspension in LB. Cultures were mixed with 10mg/mL yeast in the presence (triangles) and absence (hexagons) of 3% mannose in a 12-well plate, and the optical density of the wells at 600nm (OD 600 ) was measured via spectrophotometry. The strains used are SL257, SL253, SL255, SL289, and SL291. For all panels, data were obtained from three biological replicates with error bars representing the standard deviation of the mean. Statistical significance was determined by two-way ANOVA followed by Tukey’s post hoc multiple comparisons test. Asterisks (**, ***, ****) indicate P < 0.01, P < 0.001, and P < 0.0001, respectively, and ns = not significant.

Article Snippet: The resulting gene fragments were cloned into a XbaI/HindIII-digested pBAD24 vector ( ) using T4 DNA ligase (NEB, Ipswich, MA).

Techniques: Over Expression, Agglutination, Plasmid Preparation, Incubation, Centrifugation, Spectrophotometry, Standard Deviation

Journal: Scientific Reports

Article Title: The carRS-ompV-virK operon of Vibrio cholerae senses antimicrobial peptides and activates the expression of multiple resistance systems

doi: 10.1038/s41598-025-98217-3

Figure Lengend Snippet: Minimal inhibitory concentrations (MICs) of polymyxin B and LL-37.

Article Snippet: The amplicons and purified pBAD24 vector were digested with PstI and KpnI from New England Biolabs ® according to the manufacturer’s instructions, and purified from agarose gel using Monarch Gel purification kit (New England Biolabs ® ).

Techniques:

Journal: Scientific Reports

Article Title: The carRS-ompV-virK operon of Vibrio cholerae senses antimicrobial peptides and activates the expression of multiple resistance systems

doi: 10.1038/s41598-025-98217-3

Figure Lengend Snippet: Bacterial strains and plasmids used in this study.

Article Snippet: The amplicons and purified pBAD24 vector were digested with PstI and KpnI from New England Biolabs ® according to the manufacturer’s instructions, and purified from agarose gel using Monarch Gel purification kit (New England Biolabs ® ).

Techniques: Isolation, Infection, Derivative Assay, Expressing, Plasmid Preparation

Journal: Scientific Reports

Article Title: The carRS-ompV-virK operon of Vibrio cholerae senses antimicrobial peptides and activates the expression of multiple resistance systems

doi: 10.1038/s41598-025-98217-3

Figure Lengend Snippet: Primers used in this study.

Article Snippet: The amplicons and purified pBAD24 vector were digested with PstI and KpnI from New England Biolabs ® according to the manufacturer’s instructions, and purified from agarose gel using Monarch Gel purification kit (New England Biolabs ® ).

Techniques: Cloning