Chloramphenicol cm

Manufactured by Merck Group

Sourced in United States, Germany

Chloramphenicol (Cm) is a broad-spectrum antibiotic that inhibits bacterial protein synthesis. It is commonly used in research and laboratory settings.

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Lab products found in correlation

Ampicillin amp, by Merck Group (14 mentions) Kanamycin kan, by Merck Group (6 mentions) Kanamycin kan, by GoldBio (5 mentions) Erythromycin em, by Merck Group (3 mentions) Kanamycin kn, by Merck Group (3 mentions) L arabinose, by Merck Group (3 mentions) Agar a, by Bio Basic (3 mentions) Polymyxin b, by Merck Group (3 mentions) Cefoxitin cfx, by Merck Group (2 mentions) Yeast extract, by Thermo Fisher Scientific (2 mentions) Tetracycline tet, by Merck Group (2 mentions) Ampicillin, by Merck Group (2 mentions) Ampicillin ap, by Merck Group (2 mentions) Ciprofloxacin, by Merck Group (2 mentions) Anhydrotetracycline atc, by Takara Bio (2 mentions) Activated charcoal, by Merck Group (1 mentions) Skimmed milk, by Scharlab (1 mentions) Blood agar base bab, by Thermo Fisher Scientific (1 mentions) Diaminopimelic acid dap, by Merck Group (1 mentions) Kanamycin km, by Merck Group (1 mentions)

37 protocols using chloramphenicol cm

Brucella suis bv2 CITA 198 Cultivation

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The bacterial strains and plasmids used are listed in Additional file 1. We used B. suis bv2 CITA 198 (herein Bs2WT) because, although Bs2WT and the B. suis bv2 reference strain (B. suis bv2 Thomsen) have the same PCR-RFLP pattern [3 (link)], the former shows a virulence pattern in mice typical of B. suis bv2 field strains and the latter is attenuated (Additional file 2).
Bacteria were routinely grown either in standard tryptic soy broth (TSB; Scharlau, Barcelona, Spain) or TSA (TSB supplemented with agar [Pronadisa, Laboratorios Conda, Spain]) at 37 °C. For the studies in mice, vaccines and challenge strains were grown on Blood Agar Base (BAB; Oxoid, UK). When needed, media were supplemented with 5% sucrose, diaminopimelic acid (DAP [Sigma]; 1 mM), 0.2% activated charcoal, kanamycin (Km) at 50 µg/mL or at 35 µg/mL, ampicillin (Amp) at 100 µg/mL and/or chloramphenicol (Cm) at 20 µg/mL (all from Sigma). The lactate-glutamate-glycerol-vitamins synthetic medium of Gerhardt’s [35 (link)] was supplemented with 1 mM methionine (mGSM) (this amino acid is required for growth of some Brucella strains in synthetic media [32 (link)] including Bs2WT (Zúñiga-Ripa, unpublished observations). All strains were stored at − 80 °C in skimmed milk (Scharlau, Barcelona, Spain) or in TSB supplemented with 0.5% yeast extract (Pronadisa, Laboratorios Conda, Spain) (TYSB) and 7% dimethylsulfoxide.

Aragón-Aranda B., de Miguel M.J., Lázaro-Antón L., Salvador-Bescós M., Zúñiga-Ripa A., Moriyón I., Iriarte M., Muñoz P.M, & Conde-Álvarez R. (2020). Development of attenuated live vaccine candidates against swine brucellosis in a non-zoonotic B. suis biovar 2 background. Veterinary Research, 51, 92.

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Cultivation and Antibiotic Conditions for Bacterial Strains

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The bacterial strains and plasmids used in this study are listed in Table 1. R. anatipestifer strains were grown at 37°C in tryptic soybean broth (TSB, Oxoid) or tryptic soy agar (TSA, Oxoid) in an atmosphere of 5% CO₂. Escherichia coli (E. coli) strains were grown on Luria-Bertani (LB, Oxoid) broth or agar at 37°C. When required, antibiotics were added at the following final concentrations (μg/ml): Chloramphenicol (Cm, Sigma), 25; cefoxitin (Cfx, Sigma), 1; kanamycin (Kan, Sigma), 100; ampicillin (Amp, Sigma), 100 or spectinomycin (Spc, Sigma), 70. Diaminopimelic acid (DAP, 50 μg/ml) to E. coli X7213λpir cultures (Edwards et al., 1998 (link)).

Huang L., Yuan H., Liu M.F., Zhao X.X., Wang M.S., Jia R.Y., Chen S., Sun K.F., Yang Q., Wu Y., Chen X.Y., Cheng A.C, & Zhu D.K. (2017). Type B Chloramphenicol Acetyltransferases Are Responsible for Chloramphenicol Resistance in Riemerella anatipestifer, China. Frontiers in Microbiology, 8, 297.

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Bacterial Strain Interactions with Tetrahymena

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The bacterial strains and plasmids used in this study are listed in Table S1. A. hydrophila NJ-35 and its derivative non-Tetrahymena-exposed strain B1 (conventionally cultured without T. thermophila) and Tetrahymena-exposed strain SCV1 (a small colony variant (SCV) strain after cocultured with T. thermophila) were maintained in Luria-Bertani (LB) medium at 28°C, as previously described [12 (link)]. Escherichia coli SM10 was conventionally cultured in LB medium at 37°C [12 (link)]. When necessary, chloramphenicol (Cm) (Sigma Louis, MO, USA), kanamycin (Kan) (Sigma), or ampicillin (Amp) (Sigma) were added to the medium. T. thermophila SB210 (accession number GCA_000261185.1) was obtained from Dr. Miao Wei, Institute of Hydrobiology, China Academy of Sciences, and cultured in SPP medium (2% protease peptone, 0.1% yeast extract, 0.2% glucose, 0.003% EDTA-Fe) at 28°C [12 (link)]. The lytic bacteriophage G65, belonging to the family Myoviridae, was isolated from a contaminated river in Nanjing, China, in 2014 [20 (link)].

Dong Y., Liu J., Nie M., Zhao D., Huang H., Geng J., Wan X., Lu C, & Liu Y. (2022). Comparative transcriptome combined with morphophysiological analyses revealed the molecular mechanism underlying Tetrahymena thermophila predation-induced antiphage defense in Aeromonas hydrophila. Virulence, 13(1), 1650-1665.

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Culturing Pneumococcus and E. coli

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S. pneumoniae D39 wild type (WT) was cultured in THY medium consisting of Todd-Hewitt broth (Oxoid, United Kingdom) supplemented with 0.5% yeast extract (Oxoid, United Kingdom) or grown on Columbia agar (Difco, United States) containing 5% sheep blood (Ruite, China) in an incubator containing 5% CO₂ at 37°C. Escherichia coli BL21 was cultured in Luria-Bertani (LB) medium in a shaking incubator at 37°C. The transformed strains were selected for growth in the presence of 4 μg/ml of chloramphenicol (Cm; Sigma, United States), or 100 μg/ml of ampicillin (Amp; Sigma, United States) were added to the medium.
The iron-restricted medium was prepared by adding 5% Chelex-100 resin (Bio-Rad, United States) to THY with continuous stirring for 8 h, then filtered to remove the resin, sterilized, and supplemented with 100 μM of CaCl₂ and 2 mM of MgCl₂. If necessary, an iron source, such as hemin or FeCl₃, was added to the medium. All the bacterial strains and plasmids used in this study are listed in Table 1.

Cao L., Li N., Dong Y., Yang X.Y., Liu J., He Q.Y., Ge R, & Sun X. (2022). SPD_0090 Negatively Contributes to Virulence of Streptococcus pneumoniae. Frontiers in Microbiology, 13, 896896.

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Characterization of A. hydrophila NJ-35 Strain

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The bacterial strains and plasmids that were used in this study are listed in Table 3. A. hydrophila NJ-35, which belongs to the ST251 clonal group, was isolated from diseased cultured crucian carp in the Jiangsu province of China in 2010 [40 (link)]. The genome sequence of A. hydrophila NJ-35 has been published in GenBank (accession number CP006870). A. hydrophila and E. coli were cultured in Luria Bertani broth (LB) at 28 and 37 °C, respectively. When necessary, chloramphenicol (Cm) (Sigma Louis, MO, USA), kanamycin (Kan) (Sigma), or ampicillin (Amp) (Sigma) were added to the medium.

Dong Y., Wang Y., Liu J., Ma S., Awan F., Lu C, & Liu Y. (2018). Discovery of lahS as a Global Regulator of Environmental Adaptation and Virulence in Aeromonas hydrophila. International Journal of Molecular Sciences, 19(9), 2709.

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Culturing Bacterial Strains for Research

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The S. pneumoniae D39 was cultured in THY medium (Todd-Hewitt broth; Oxoid, UK) with 0.5% yeast extract (Oxoid) or on Columbia agar (Difco, USA) containing 5% sheep blood (Ruite, China) at 37°C in a 5% CO₂ incubator (MCO-170AICUVHL-PC; Panasonic, Japan). Escherichia coli DH5ɑ and BL21 (Invitrogen, USA) were both cultured in Luria-Bertani (LB) medium at 37°C in an incubator shaker (Yi Heng, China). All strains were stored at –80°C in 15% glycerol. Erythromycin (Erm; Sigma, USA) at 0.25 mg/ml, chloramphenicol (Cm; Sigma) at 4 mg/ml, or ampicillin (Amp; Sigma) at 100 μg/ml was added to the medium when needed for selective culture. All of the bacterial strains and plasmids used are listed in Table 3.

Zheng Y.D., Pan Y., He K., Li N., Yang D., Du G.F., Ge R., He Q.Y, & Sun X. (2020). SPD_1495 Contributes to Capsular Polysaccharide Synthesis and Virulence in Streptococcus pneumoniae. mSystems, 5(1), e00025-20.

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Bacterial Propagation and Strain Selection

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The bacterial strains, plasmids, and primers used in this study are listed in Table 1. Strains of L. acidophilus were propagated statically in de Man-Rogosa-Sharpe (MRS) broth (Difco Laboratories, Inc., Detroit, MI) or on MRS agar (1.5% [wt/vol]; Difco) under aerobic conditions at 37°C or 42°C. Transformants were selected in the presence of 2 μg/ml erythromycin (Em) (Sigma-Aldrich, St. Louis, MO) and/or 2 to 5 μg/ml chloramphenicol (Cm) (Sigma). Escherichia coli was grown in brain heart infusion (BHI) medium (Difco) at 37°C with shaking aeration. E. coli EC101 was propagated in the presence of 40 μg/ml kanamycin (Km) (39 (link)). For upp-based counterselective gene replacement procedures, plasmid-free double recombinants were selected on a glucose semidefined agar medium containing 100 μg/ml 5-fluorouracil (5-FU) (Sigma), as previously described (38 (link)).

Hymes J.P., Johnson B.R., Barrangou R, & Klaenhammer T.R. (2016). Functional Analysis of an S-Layer-Associated Fibronectin-Binding Protein in Lactobacillus acidophilus NCFM. Applied and Environmental Microbiology, 82(9), 2676-2685.

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Salmonella Typhimurium Strain Maintenance

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S. Typhimurium strains and plasmids used in this work are listed in S1 Table. Strains were maintained in Luria-Bertani (LB) broth or on plates with appropriate antibiotics at the following concentrations; ampicillin (Sigma Aldrich), 100 μg.ml^-1; chloramphenicol (Cm, Sigma Aldrich), 12.5 μg.ml^-1; kanamycin (Kn, Sigma Aldrich), 50 μg.ml^-1; tetracycline (Tet, Sigma Aldrich), 15 μg.ml^-1. M9 minimal medium with 0.4% w/v glucose was used where indicated. Oligonucleotide primers were purchased from Sigma Genosys or Illumina.

Garcia-Gutierrez E., Chidlaw A.C., Le Gall G., Bowden S.D., Tedin K., Kelly D.J, & Thompson A. (2016). A Comparison of the ATP Generating Pathways Used by S. Typhimurium to Fuel Replication within Human and Murine Macrophage and Epithelial Cell Lines. PLoS ONE, 11(3), e0150687.

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Cultivation and Preservation of L. acidophilus

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All bacterial strains and plasmids are listed in Table 1. Bacterial cultures were cryopreserved in their respective media with a 15% glycerol concentration (vol/vol) and stored at -80°C. L. acidophilus was propagated in de Mann, Rogosa and Sharpe (MRS) (Difco Laboratories, Inc., Detroit, MI, USA) broth under static aerobic conditions at 37°C, or on MRS agar (1.5% wt/vol agar, Difco) incubated anaerobically at 37°C for 48 h. Concentrations of 2 μg/mL of erythromycin (Em) (Sigma-Aldrich, St. Louis, MO, USA) and 2–5 μg/mL of chloramphenicol (Cm) (Sigma) were used for plasmid selection in L. acidophilus NCFM, when appropriate. Selection for 5-fluorouracil resistant L. acidophilus was performed by supplementing glucose semi-defined (GSDM) (Kimmel and Roberts, 1998 (link)) agar with a final concentration of 100 μg/mL of 5- fluorouracil (Sigma) (Goh et al., 2009 (link)). Escherichia coli EC1000 was propagated aerobically in Luria-Bertani (Difco) broth at 37°C, or on brain-heart infusion (Difco) solid medium supplemented with 1.5% agar. Antibiotic selection of E. coli was maintained with 40 μg/mL kanamycin (Kn) and 150 μg/mL of Em for recombinant E. coli, when appropriate.

Selle K., Goh Y.J., Johnson B.R., O’Flaherty S., Andersen J.M., Barrangou R, & Klaenhammer T.R. (2017). Deletion of Lipoteichoic Acid Synthase Impacts Expression of Genes Encoding Cell Surface Proteins in Lactobacillus acidophilus. Frontiers in Microbiology, 8, 553.

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Propagation and Storage of Bacterial Strains

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Example 1

All bacterial strains are listed in Table 1. Bacterial cultures were cryopreserved in an appropriate growth medium with 25% glycerol (vol/vol) and stored at −80° C. S. thermophilus was propagated in Elliker media (Difco) supplemented with 1% beef extract (wt/vol) and 1.9% (wt/vol) β-glycerolphosphate (Sigma) broth under static aerobic conditions at 37° C., or 10 on solid medium with 1.5% (wt/vol) agar (Difco), incubated anaerobically at 37° C. for 48 hours. Concentrations of 2 pg/mL of erythromycin (Em) and 5 μg/mL of chloramphenicol (Cm) (Sigma) were used for plasmid selection in S. thermophilus, when appropriate. E. coli EC1000 was propagated aerobically in Luria-Bertani (Difco) broth at 37° C., or on brain-heart infusion (BHI) (Difco) solid medium supplemented with 1.5% agar. Antibiotic selection of E. coli was maintained with 40 μg/mL kanamycin (Kn) and 150 μg/mL of Em for recombinant E. coli, when appropriate. Screening of S. thermophilus derivatives for β-galactosidase activity was assessed qualitatively by supplementing a synthetic Elliker medium with 1% lactose, 1.5% agar, and 0.04% bromo-cresol purple as a pH indicator.

US10136649B2. Methods for screening bacteria, archaea, algae, and yeast using CRISPR nucleic acids (2018-11-27). North Carolina State University [US]. Inventors: Rodolphe Barrangou [US], Kurt M. Selle [US].

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