psam arac  (Addgene inc)

Journal: Communications Biology

Article Title: Genome-wide identification of bacterial colonization and fitness determinants on the floating macrophyte, duckweed

doi: 10.1038/s42003-022-03014-7

Figure Lengend Snippet: a Schematic diagram of the screening experiments. The transposon mutant library of Aquitalea magnusonii H3rifR was inoculated to flasks with and without duckweed, and the DNA of mutants was collected from three fractions after 3 h or 7 d. Samples were destructively obtained from four flasks, and pooled before DNA extraction. b The abundance of mutant cells in the three fractions during the screening experiments. Error bars show standard deviation. c Detection frequency of genes in the three fractions after 3 h and 7 d. Log 10 gene-level detection frequencies were compared. Depleted and enriched genes were determined based on the effect size (fold change > 2) and statistical significance ( q < 0.05) among Plant and Control samples. Statistical test was performed by considering different mutants (insertion sites) of the same gene as replicates. d Comparison of the screening results of 3-h and 7-d experiments. Genes showed significant depletion or enrichment in the 3-h and 7-d experiments are denoted by crosses and squares, respectively.

Article Snippet: The mutant library of H3rifR was created with transposon vector pSAM_AraC (Addgene plasmid #91569).

Techniques: Mutagenesis, DNA Extraction, Standard Deviation, Control, Comparison

Journal: Methods in Molecular Biology

Article Title: Proteus mirabilis

doi: 10.1007/978-1-4939-9601-8

Figure Lengend Snippet: Fig. 1 P. mirabilis colonies on agar. (a) Low salt LB. (b) A mix of P. mirabilis (white) and E. coli (pink) on MacConkey agar. (c, d) A mix of P. mirabilis and E. coli on blood agar. What appears to be isolated colonies in panel c is actually covered with swarming P. mirabilis, which becomes evident when a loop is drawn across the background (d). In all panels, the P. mirabilis strain is HI4320. In panels b–d, the uropathogenic E. coli strain is CFT073

Article Snippet: Escherichia coli S17-1λpir + pSAM_AraC (Addgene plasmid #91569) (see Note 2).

Techniques: Isolation

Journal: Methods in Molecular Biology

Article Title: Proteus mirabilis

doi: 10.1007/978-1-4939-9601-8

Figure Lengend Snippet: Fig. 1 Siderophore production detected by CAS shuttle solution. Siderophore production measured at 630 nm (A630) using a CAS assay and filtered bacterial supernatants following culture in MOPS defined medium with (black bars) and without (white bars) 0.1 mM FeCl3·6H2O of P. mirabilis HI4320 wild type and pbtA (proteobactin), nrpR (yersiniabactin-like), and nrpR pbtA (double) mutants and E. coli CFT073 and entF iucB (enterobactin aerobactin) double mutant. P. mirabilis supernatants were concentrated 50-fold. Gray bar, positive control of phosphate buffered saline containing 15 μM deferoxamine (Desferal). A lower A630 indicates greater siderophore production. Significant differences (∗, P 0.0022) were determined using a two-tailed unpaired t-test; bars show standard error of the mean. (From: Himpsl, SD, Pearson MM, Arewa˚ng CJ, Nusca TD, Sherman DH, Mobley HLT. 2010. Proteobactin and a yersiniabactin-related siderophore mediate iron acquisition in Proteus mirabilis. Mol Microbiol. 2010 Oct; 78(1): 138–157)

Article Snippet: Escherichia coli S17-1λpir + pSAM_AraC (Addgene plasmid #91569) (see Note 2).

Techniques: Mutagenesis, Positive Control, Saline, Two Tailed Test

Journal: Methods in Molecular Biology

Article Title: Proteus mirabilis

doi: 10.1007/978-1-4939-9601-8

Figure Lengend Snippet: Fig. 2 Siderophore production on CAS agar. Overnight cultures of E. coli CFT073 and entF iucB (enterobactin aerobactin) double mutant, and P. mirabilis HI4320 and nrpR (yersiniabactin-like), pbtA (proteobactin), and nrpR pbtA (double) mutants were grown to stationary phase in LB and spotted in 5 μL volumes onto CAS agar, left to dry, inverted, and incubated at 30 C for 18 h. A color change of chrome azurol S (CAS) agar from blue to orange indicates siderophore production. (From: Himpsl, SD, Pearson MM, Arewa˚ng CJ, Nusca TD, Sherman DH, Mobley HLT. 2010. Proteobactin and a yersiniabactin-related siderophore mediate iron acquisition in Proteus mirabilis. Mol Microbiol. 2010 Oct; 78(1): 138–157)

Article Snippet: Escherichia coli S17-1λpir + pSAM_AraC (Addgene plasmid #91569) (see Note 2).

Techniques: Mutagenesis, Incubation

Journal: Methods in Molecular Biology

Article Title: Proteus mirabilis

doi: 10.1007/978-1-4939-9601-8

Figure Lengend Snippet: Fig. 3 P. mirabilis is unable to utilize yersiniabactin produced by uropathogenic E. coli 536. LB medium was supplemented with 25 μM deferoxamine, the minimum concentration of chelator required to completely suppress growth of P. mirabilis HI4320. Lactose-fermenting E. coli were distinguished from P. mirabilis by using the chromogenic indicator X-gal. (a) Growth of wild-type P. mirabilis, the proteobactin synthesis/ receptor mutant (PbtSR), and the yersiniabactin-related siderophore/receptor mutant (NrpSR) is restored when cross-fed by E. coli 536 that produces both enterobactin and yersiniabactin. (b and c) Restoration of P. mirabilis growth for all strains tested is abolished when cross-fed by either the E. coli 536 enterobactin mutant, entF::kan, or the enterobactin/yersiniabactin double mutant, entF ybtS::kan. (d) P. mirabilis growth is restored using E. coli CFT073, an enterobactin-producing strain incapable of synthesizing yersiniabactin. (From: Himpsl SD, Pearson MM, Arewa˚ng CJ, Nusca TD, Sherman DH, Mobley HLT. 2010. Proteobactin and a yersiniabactin-related siderophore mediate iron acquisition in Proteus mirabilis. Mol Microbiol. 2010 Oct; 78 (1): 138–157)

Article Snippet: Escherichia coli S17-1λpir + pSAM_AraC (Addgene plasmid #91569) (see Note 2).

Techniques: Produced, Concentration Assay, Mutagenesis

Journal: Methods in Molecular Biology

Article Title: Proteus mirabilis

doi: 10.1007/978-1-4939-9601-8

Figure Lengend Snippet: Fig. 1 Hemagglutination results for E. coli and P. mirabilis. Bacteria were diluted twofold in PBS across ten wells of a 96-well plate (the first six columns are shown). The second to last column of the plate contains undiluted bacteria and erythrocytes with 50 mM mannose (+ man). 3% (vol/vol) guinea pig erythrocytes in PBS were used to detect hemagglutination by mannose-resistant/Proteus-like (MR/P) fimbriae in P. mirabilis and mannose-sensitive hemagglutination by type 1 fimbriae in E. coli

Article Snippet: Escherichia coli S17-1λpir + pSAM_AraC (Addgene plasmid #91569) (see Note 2).

Techniques: Bacteria