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Amsacrine

Manufactured by Abcam
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Amsacrine is an organic compound with the chemical formula C₂₂H₂₃N₃O₂. It is a topoisomerase II inhibitor and exhibits cytotoxic activity.

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Lab products found in correlation

E coli dna gyrase tg2000g, by TopoGEN (2 mentions) Dna gyrase assay kit tg1003, by TopoGEN (2 mentions) 0.2 μm syringe filter, by Corning (1 mentions) Crystal violet, by Merck Group (1 mentions) Bacto agar, by BD (1 mentions) Safranin, by Merck Group (1 mentions) Hematoxylin and eosin, by Merck Group (1 mentions) 2 methyl 2 butanol, by Merck Group (1 mentions) Trypticase soy broth tsb, by BD (1 mentions) 2 2 2 tribromoethanol, by Merck Group (1 mentions) Scfas, by Merck Group (1 mentions)

5 protocols using amsacrine

1

Transduction of S. aureus using Φ85 bacteriophage

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S. aureus was grown in tryptic soy broth (TSB) or on TSB with 1.5% agar at 30 °C. Transduction was performed using Φ85 as described40 (link). Antibiotic concentrations used were 5 μg/ml erythromycin (Erm). Amsacrine was purchased from Abcam (ab142742). E. coli DNA gyrase (TG2000G) and DNA gyrase assay kit (TG1003) was purchased from TopoGEN.
Pasquina L., Maria JP J.r., Wood B.M., Moussa S.H., Matano L., Santiago M., Martin S.E., Lee W., Meredith T.C, & Walker S. (2015). A synthetic lethal approach for compound and target identification in Staphylococcus aureus. Nature chemical biology, 12(1), 40-45.
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2

Preparation and Characterization of Short-Chain Fatty Acids

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SCFAs were purchased from Sigma-Aldrich Inc. (St. Louis, MO, United States). SCFAs were dissolved in endotoxin-free distilled water (Dai Han Pharm Co. Ltd., Seoul, South Korea) and filtered with a syringe filter (0.2 μm) purchased from Corning (Corning, NY, United States) prior to use. Luria-Bertani (LB) broth was purchased from LPS solution (Daejeon, South Korea). Trypticase soy broth (TSB) and Bacto agar were purchased from BD Biosciences (Franklin Lakes, NJ, United States). 2,2,2-Tribromoethanol and 2-methyl-2-butanol were purchased from Sigma-Aldrich Inc. Amsacrine (AMSA) was purchased from Abcam (Cambridge, United Kingdom). Hematoxylin and eosin were purchased from Sigma-Aldrich Inc. and BBC Biochemical (Mount Vernon, WA, United States), respectively. Crystal violet and safranin were purchased from Sigma-Aldrich Inc. Iodide solution was purchased from Samchun Chemicals (Seoul, South Korea).
Jeong S., Kim H.Y., Kim A.R., Yun C.H, & Han S.H. (2019). Propionate Ameliorates Staphylococcus aureus Skin Infection by Attenuating Bacterial Growth. Frontiers in Microbiology, 10, 1363.
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3

Screening S. aureus with Chemical Analogs

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S. aureus was grown in tryptic soy broth (TSB) or on TSB with 1.5% agar at 30°C. High-throughput screening was done at the ICCB-Longwood Screening Facility. DBI-1 and DBI-3 were purchased from Life Chemicals (catalog numbers F0715-0438 and F2553-0110 respectively). Analogs DBI-2, DBI-4, DBI-5, and DBI-6 were purchased from Life Chemicals (catalog numbers Z763933882, Z27852313, Z119631630 and Z27772254 respectively). Amsacrine was purchased from Abcam (ab142742).
Matano L.M., Morris H.G., Wood B.M., Meredith T.C, & Walker S. (2016). Accelerating the discovery of antibacterial compounds using pathway-directed whole cell screening. Bioorganic & medicinal chemistry, 24(24), 6307-6314.
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4

Transduction of S. aureus using Φ85 bacteriophage

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S. aureus was grown in tryptic soy broth (TSB) or on TSB with 1.5% agar at 30 °C. Transduction was performed using Φ85 as described40 (link). Antibiotic concentrations used were 5 μg/ml erythromycin (Erm). Amsacrine was purchased from Abcam (ab142742). E. coli DNA gyrase (TG2000G) and DNA gyrase assay kit (TG1003) was purchased from TopoGEN.
Pasquina L., Maria JP J.r., Wood B.M., Moussa S.H., Matano L., Santiago M., Martin S.E., Lee W., Meredith T.C, & Walker S. (2015). A synthetic lethal approach for compound and target identification in Staphylococcus aureus. Nature chemical biology, 12(1), 40-45.
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5

Microbial Transduction and Protein Expression

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Strains are listed in Supplementary Table 5. S. aureus was grown in tryptic soy broth (TSB) or on TSB with 1.5% agar at 30 and 37 °C. Phage 80alpha was used for transductions and the methods for transduction were described previously40 (link). All constructed LCP-pET28-plasmids were transformed into NovaBlue(DE3) E. Coli, and LCP proteins were overexpressed in BL21(DE3) E. Coli grown in lysis broth (LB). Antibiotics were used at the following concentrations: 10 µg ml−1 chloramphenicol, 10 µg ml−1 erthromycin, 50 µg ml−1 kanamycin, 50 µg ml−1 neomycin, 50 µg ml−1 carbenicillin. L-[14C]-Glycerol-3-phosphate was purchased from Person-Elmer. UDP-[14C]-GlcNAc was purchased from American Radiolabeled Chemicals, Inc (ARC). Amsacrine was purchased from Abcam (ab142742).
Schaefer K., Matano L.M., Qiao Y., Kahne D, & Walker S. (2017). In vitro reconstitution demonstrates the cell wall ligase activity of LCP proteins. Nature chemical biology, 13(4), 396-401.
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