Journal: International Journal of Molecular Sciences

Article Title: Lon Protease Is Important for Growth under Stressful Conditions and Pathogenicity of the Phytopathogen, Bacterium Dickeya solani

doi: 10.3390/ijms21103687

Figure Lengend Snippet: Confirmation of successful deletion of the lon gene in D. solani IPO 2222: ( A ) PCR analysis of genomic DNA isolated from the D. solani WT (wild type) and Δ lon mutant. The lon gene was replaced by a 1000 bp smaller kanamycin resistance gene. Δlon1 and Δlon2 denote two independent clones ( B ) immunodetection using the anti-Lon E. coli primary antibodies. ( C ) The qPCR analysis with the use of the lon gene-specific primers revealed that no cDNA amplification product was created within 50 cycles.

Article Snippet: Briefly, the Lon protein was detected with the anti- Escherichia coli Lon rabbit antibodies (#40219-T24, Sino Biological Inc., Beijing, China) at dilution 1:2000 followed by incubation with HRP conjugated secondary anti-rabbit antibodies (#31462 Thermo Fisher) diluted 1:50,000.

Techniques: Isolation, Mutagenesis, Clone Assay, Immunodetection, Amplification

Journal: International Journal of Molecular Sciences

Article Title: Lon Protease Is Important for Growth under Stressful Conditions and Pathogenicity of the Phytopathogen, Bacterium Dickeya solani

doi: 10.3390/ijms21103687

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

Article Snippet: Briefly, the Lon protein was detected with the anti- Escherichia coli Lon rabbit antibodies (#40219-T24, Sino Biological Inc., Beijing, China) at dilution 1:2000 followed by incubation with HRP conjugated secondary anti-rabbit antibodies (#31462 Thermo Fisher) diluted 1:50,000.

Techniques:

Journal: EBioMedicine

Article Title: Gut bacterial type III secretion systems aggravate colitis in mice and serve as biomarkers of Crohn's disease.

doi: 10.1016/j.ebiom.2024.105296

Figure Lengend Snippet: Fig. 1: A. pulmonis mAT1 harbors a functional T3SS. (A) Phylogenetic tree of A. pulmonis mAT1 (red), compared to A. dolens LMG 26841 strain and 14 closest type strains automatically picked by the Type Strain Genome Server (TYGS). The phylogenetic tree was generated by TYGS with default settings. (B) 35 online-available Achromobacter genomes were re-annotated by NR and searched for genes of T3SS proteins that belong to each T3SS ortholog. The quantities of each T3SS ortholog were plotted in the heatmap. Achromobacter strains were classified into host- isolated and environmental-isolated groups. Achromobacter strains in each group were organized by phylogenic tree generated by GTDB-Tk and IQ-TREE2 software. Achromobacter strains that contain more than 10 T3SS orthologs annotated by NR are considered T3SS-harboring strains and labeled red. (C) Neighbor-joining phylogenetic tree T3SS ATPase SctN of representative T3SS-harboring bacteria strains. Bacteria names are color-coded to indicate designated T3SS families. (D) T3SS gene cluster of A. pulmonis mAT1 compared with that of B. bronchiseptica RB50. Genes are color-coded to indicate designated components of the T3SS apparatus. (E) The relative expression level of T3SS genes of A. pulmonis mAT1 grown in designated media, measured by qRT-PCR. All data are expressed as the mean ± SEM of at least three experimental repeats, each with two technical replicates. The Kruskal–Wallis test with Dunne’s multiple comparisons was performed and adjusted by the Benjamini-Hochberg method. For Dunn’s multiple comparisons test, the mean of each group was compared with the mean of the control group. *p < 0.05, **p < 0.01, ***p < 0.001. (F) Immunoblotting of secreted AopD by designated strains. Each bacterial culture was centrifuged

Article Snippet: The proteins in the supernatant fraction were precipitated in 10% (v/v) trichloroacetic acid.26 Proteins in both fractions were separated via 12% SDS-PAGE and analyzed by immunoblotting with a rabbit anti-AopD polyclonal antibody and a rabbit anti-DnaK polyclonal antibody (CUSABIO; #CSB-PA633459HA01EGW).

Techniques: Functional Assay, Generated, Isolation, Software, Labeling, Bacteria, Expressing, Quantitative RT-PCR, Control, Western Blot