Journal: The Journal of Biological Chemistry

Article Title: Der f 34, a Novel Major House Dust Mite Allergen Belonging to a Highly Conserved Rid/YjgF/YER057c/UK114 Family of Imine Deaminases

doi: 10.1074/jbc.M116.728006

Figure Lengend Snippet: Imine deaminase activity of Der f 34. Imine deaminase activity of Der f 34 was analyzed using rTF-Der f 34, recombinant RidA protein from S. enterica as a positive control, mutant rTF-Der f 34, nonfunctional variant RidA protein (RidAR105A), rTF as a negative control, and Leu, Met, or Phe as a substrate. In the assay, l-amino acid oxidase dehydrated an amino acid to produce an imine, and the resultant imine reacted with semicarbazide to produce semicarbazone that has an absorbance at 248 nm. An imine deaminase accelerates the hydrolysis of substrate-derived imine intermediates to prevent the production of semicarbazone (26, 37). Semicarbazone formation at each time point from 0 min for vehicle (buffer, black diamonds), rTF-Der f 34 (red diamonds), mutant rTF-Der f 34 (purple diamonds), RidA protein (orange diamonds), variant RidAR105A protein (green diamonds), and rTF (bright blue diamonds) is plotted on the y axis, and the incubation time for the reaction is shown on the x axis. The data represent the average ± S.D. (error bars) for four independent experiments. Non-repeated ANOVA was applied to compare data among the groups and, when the differences were significant (p < 0.01), post hoc analysis using an SNK test was performed. Significant differences as estimated by post hoc SNK test compared with rTF or vehicle as a negative control are shown as follows: p < 0.01 for rTF (**) and p < 0.05 (#) or p < 0.01 (##) for vehicle, respectively.

Article Snippet: Briefly, 300 n m rTF-Der f 34, mutant rTF-Der f 34, wild type RidA protein from S. enterica (NCBI GenBank TM accession number {"type":"entrez-nucleotide","attrs":{"text":"AF095578","term_id":"4558863","term_text":"AF095578"}} AF095578 ), inactive variant RidA R105A protein, or rTF as a negative control was mixed with 10 m m semicarbazide-HCl, 1 μg (2–5 units) of bovine liver catalase, 10 μg (30 milliunits) of l -amino acid oxidase from Crotalus adamanteus (Sigma-Aldrich), and substrate amino acid: 5 m m Leu, 6.25 m m Met, or 10 m m Phe in 50 m m potassium pyrophosphate (pH 8.7) ( 26 , 37 ).

Techniques: Activity Assay, Recombinant, Positive Control, Mutagenesis, Variant Assay, Negative Control, Derivative Assay, Incubation

Journal: International Journal of Microbiology

Article Title: Distribution and Genetic Diversity of Salmonella enterica in the Upper Suwannee River

doi: 10.1155/2011/461321

Figure Lengend Snippet: DiversiLab rep-PCR dendrogram. Representative strains are shown to demonstrate the segregation of different serotypes by rep-PCR. Cluster A was comprised of duplicate serovars of subspecies I (Typhimurium, Choleraesuis, and Enteritidis), while cluster B included duplicate serovars of subspecies III (Arizonae). Within each individual serovar, analyzed in triplicates, >94% DNA similarity was observed. More than 70% DNA relatedness was noted between serovars of the same serovars. The scale at the bottom indicates the percentage similarity among fingerprints of each genotype.

Article Snippet: PCR amplification used S. enterica rep-PCR DNA Fingerprinting Kit (BioMerieux) with AmpliTaq DNA polymerase and GeneAmp 10x PCR Buffer I with Mg Cl 2 (Applied Biosystems, Inc.).

Techniques:

Journal: International Journal of Microbiology

Article Title: Distribution and Genetic Diversity of Salmonella enterica in the Upper Suwannee River

doi: 10.1155/2011/461321

Figure Lengend Snippet: Distribution of rep-PCR genogroups related to the source of Salmonella isolates.

Article Snippet: PCR amplification used S. enterica rep-PCR DNA Fingerprinting Kit (BioMerieux) with AmpliTaq DNA polymerase and GeneAmp 10x PCR Buffer I with Mg Cl 2 (Applied Biosystems, Inc.).

Techniques:

Journal: International Journal of Microbiology

Article Title: Distribution and Genetic Diversity of Salmonella enterica in the Upper Suwannee River

doi: 10.1155/2011/461321

Figure Lengend Snippet: Distribution of S. enterica genogroups by location for strains recovered from the Suwannee River.

Article Snippet: PCR amplification used S. enterica rep-PCR DNA Fingerprinting Kit (BioMerieux) with AmpliTaq DNA polymerase and GeneAmp 10x PCR Buffer I with Mg Cl 2 (Applied Biosystems, Inc.).

Techniques: Sampling

Journal: International Journal of Microbiology

Article Title: Distribution and Genetic Diversity of Salmonella enterica in the Upper Suwannee River

doi: 10.1155/2011/461321

Figure Lengend Snippet: Distribution of S. enterica genogroups by month for strains recovered from the Suwannee River.

Article Snippet: PCR amplification used S. enterica rep-PCR DNA Fingerprinting Kit (BioMerieux) with AmpliTaq DNA polymerase and GeneAmp 10x PCR Buffer I with Mg Cl 2 (Applied Biosystems, Inc.).

Techniques:

Journal: International Journal of Microbiology

Article Title: Distribution and Genetic Diversity of Salmonella enterica in the Upper Suwannee River

doi: 10.1155/2011/461321

Figure Lengend Snippet: Detailed serology of S. enterica isolated from Suwannee River water.

Article Snippet: PCR amplification used S. enterica rep-PCR DNA Fingerprinting Kit (BioMerieux) with AmpliTaq DNA polymerase and GeneAmp 10x PCR Buffer I with Mg Cl 2 (Applied Biosystems, Inc.).

Techniques: Isolation