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Escherichia coli k12

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Escherichia coli K12 is a non-pathogenic strain of the bacterium Escherichia coli. It is a commonly used model organism in microbiology and molecular biology research.

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Lab products found in correlation

Anhydrous sodium carbonate, by Merck Group (1 mentions) Staphylococcus aureus, by American Type Culture Collection (1 mentions) Folin ciocalteu s phenol, by Merck Group (1 mentions) Microbiological agar, by Thermo Fisher Scientific (1 mentions) Raw 264.7 cells, by American Type Culture Collection (1 mentions) Mycobacterium smegmatis, by American Type Culture Collection (1 mentions) Mycobacterium tuberculosis h37rv, by BEI Resources (1 mentions) Twistamp basic kit, by Twist Bioscience (1 mentions) Bacillus subtilis, by American Type Culture Collection (1 mentions) Q5 high fidelity pcr kit, by New England Biolabs (1 mentions)

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Escherichia coli (416 citations) Escherichia coli K12 MG1655 (3 citations) Escherichia coli K12 ATCC10798 (2 citations) Escherichia coli (strain K12, (2 citations)

12 protocols using escherichia coli k12

1

Chitosan-Zein Antimicrobial Film Development

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Chitosan (Êxodo Científica, Sumaré, SP, Brazil), glycerol, 2,2-diphenyl-1-picrylhydrazyl (DPPH), Folin Ciocalteu’s phenol, anhydrous sodium carbonate, zein (Sigma Aldrich, St. Louis, MO, USA), and acetic acid were of analytical grade. Water (Milli-Q), acetonitrile, and standards of cyanidin-3-glucoside (Cn-3-Glu), caffeic acid, quercetin, Kaempferol, trans-ferulic acid, trans-cinnamic acid, p-coumaric acid, caffeic acid, and gallic acid were of HPLC grade. For the antimicrobial test, Nutrient, Müller-Hinton broth was used. All reagents were purchased from Sigma Aldrich (St. Louis, MO, USA). The bacteria strain used in the antimicrobial test were Escherichia coli K12 (ATCC 10798) and Staphylococcus aureus (ATCC 12598), gained from Cederlane (Burlington, CA, USA).
Avila L.B., Pinto D., Silva L.F., de Farias B.S., Moraes C.C., Da Rosa G.S, & Dotto G.L. (2022). Antimicrobial Bilayer Film Based on Chitosan/Electrospun Zein Fiber Loaded with Jaboticaba Peel Extract for Food Packaging Applications. Polymers, 14(24), 5457.
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2

Haemophilus ducreyi Antibiotic Resistance Strains

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Nine strains of Haemophilus ducreyi were chosen to reflect the varying degrees of antibiotic resistance found clinically: 35000HP, CIP542, HMC46 (PcR, TcR, CmR), HMC48 (PcR, TcR, CmR), HMC56 (PcR, TcR), HMC88 (PcR, TcR, CmR, TMPR, StrR, KanR), HMC49, HMC53 (PcR, TcR, CmR) and HMC112
[10 (link)]. In addition, Escherichia coli K-12, Pseudomonas aeruginosa 49189 (ATCC, Manassa, VA, USA), Lactobacillus reuteri HM-102 (BEI Resources, Manassas, VA, USA), and Staphylococcus aureus strain +4651 (Presque Isle Cultures, Erie, PA, USA) were used as controls. All strains of H. ducreyi were provided courtesy of Stanley Spinola (Indiana University School of Medicine) or Patricia Totten (University of Washington). S. aureus and E. coli were chosen as positive controls because all selected essential oils are effective bacteriostatic agents against both
[9 (link)]. P. aeruginosa was chosen as a control representing relative resistance to essential oils, being resistant to most essential oils except cinnamon and oregano
[9 (link)]. L. reuteri was chosen as a representative of the many Lactobacillus species that make up the vaginal flora. The susceptibility of L. reuteri is unknown, although Lactobacillus strains are resistant to tea tree oil concentrations up to 2.0%
[11 (link)].
Lindeman Z., Waggoner M., Batdorff A, & Humphreys T.L. (2014). Assessing the antibiotic potential of essential oils against Haemophilus ducreyi. BMC Complementary and Alternative Medicine, 14, 172.
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3

Bacterial and Fungal Cell Cultivation

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We used a methicillin-resistant Staphylococcus aureus (MRSA), CA-MRSA strain USA300 LAC (Los Angeles County clone), Escherichia coli K12 (ATCC, Manassass VA) and Candida albicans DAY286 reference strain (a gift from Aaron Mitchell, Department of Microbiology, Columbia University, New York, NY). Bacterial cells were grown in brain-heart infusion (BHI) medium at 37°C and Candida cells were grown in yeast extract peptone dextrose (YPD) media at 30°C. Cells were grown overnight to stationary phase and refreshed for 2 hours for bacteria and 4 hours for Candida the next day to mid-log phase. Cells were collected by centrifugation at 3500 pm for 10 minutes and resuspended in phosphate buffer at a density of 10(8) cells/mL for bacteria and 10(7) cells/mL for Candida for further experiments. Cell numbers were estimated by measuring the optical density [OD] at 600 nm (OD of 0.5=10(8) cells/mL). To enumerate CFU/mL a 10 μl aliquot of cells was serially diluted 10-fold in PBS to give dilutions of 10−1 to 10−5 times in addition to the original concentration, and 10μL aliquots of each dilution were streaked horizontally on square BHI (bacteria) or YPD (Candida) agar plates. Plates were streaked in triplicate and incubated for 18 hours at 30°C (Candida) or 37°C in the dark to allow colony formation.
Wu X., Huang Y.Y., Kushida Y., Bhayana B, & Hamblin M.R. (2016). Broad-spectrum antimicrobial photocatalysis mediated by titanium dioxide and UVA is potentiated by addition of bromide ion via formation of hypobromite. Free radical biology & medicine, 95, 74-81.
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4

Cultivation of S. aureus and E. coli

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Staphylococcus aureus 8325-4 and Escherichia coli K-12 (ATCC, Manassas, VA) were employed. Planktonic bacteria cells were cultured in brain heart infusion broth (BHI) (Fisher Scientific) with shaking at 37°C. PBS Response was used to wash microbial cells and for serial dilution. Liquid growth media was prepared from 200 ml of distilled water and 6 g of BHI powder. All liquid media were autoclaved at 120° C for 15 min before use. Solid growth media was prepared from liquid growth media with the addition of 1,5% microbiological agar (Fisher Scientific). A suspension of microbial cells was prepared by refreshing an overnight stationary phase culture in fresh medium for about 1 h. Cell pellets were isolated by centrifugation (13,690 × g for 5 min) and resuspended the appropriate volume of sterile PBS to give the desired cell-density (OD at 600 nm equivalent to 108 CFU/ml).
Kasimova K.R., Sadasivam M., Landi G., Sarna T, & Hamblin M.R. (2014). Potentiation of photoinactivation of Gram-positive and Gram-negative bacteria mediated by six phenothiazinium dyes by addition of azide ion. Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology, 13(11), 1541-1548.
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5

Viral and Bacterial Propagation Protocols

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The cytopathogenic HAV strain HM-175 (ATCC VR-1402), the murine norovirus strain MNV S99 used as proxy for hNoV and provided courtesy of the Friedrich-Loeffler-Institut in Germany (Mueller 2007) were propagated, assayed and titrated on FRhK-4 cells (ATTC CRL1688) and RAW 264.7 cells (ATCC TIB-71), respectively, as described previously (Butot et al. 2018 (link)). Viral stock titers were 6.6 and 7.6 log10 50% tissue culture infective dose (TCID50)/mL for HAV and MNV, respectively. MS2 bacteriophage (ATCC 15597-B1) was propagated, assayed and enumerated in Escherichia coli K12 (ATCC 23631), as described previously (Butot et al. 2018 (link)). The Qβ bacteriophage (ATTC 23631-B1) was propagated, assayed and enumerated using the same protocols as for MS2. The MS2 and Qβ stock concentrations were 11.3 and 10.1 log10 plaque forming units (PFU)/mL, respectively.
Butot S., Galbusera L., Putallaz T, & Zuber S. (2021). Electron Beam Susceptibility of Enteric Viruses and Surrogate Organisms on Fruit, Seed and Spice Matrices. Food and Environmental Virology, 13(2), 218-228.
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6

Culturing Diverse Bacterial Strains

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Mycobacterium tuberculosis (H37Rv) was obtained through BEI Resources, NIAID/NIH. Mycobacterium smegmatis (ATCC 607), Staphylococcus aureus (ATCC BAA-1556), and Escherichia coli K-12 (ATCC 29425) were obtained from ATCC. A single colony of bacterial strain was obtained on a Difco Middlebrook 7H10 nutrient agar enriched with 10% oleic acid, albumin, dextrose and catalase (OADC for M. tuberculosis), albumin, dextrose and catalase (ADC for M. smegmatis), and on Tryptic Soy agar (for E. coli and S. aureus). Seed cultures were obtained in Middlebrook 7H9 broth enriched with OADC (for M. tuberculosis), ADC (for M. smegmatis), and in Tryptic Soy broth (for E. coli and S. aureus), respectively. Each bacterium was grown to mid-log phase.
Siricilla S., Mitachi K., Skorupinska-Tudek K., Swiezewska E, & Kurosu M. (2014). Biosynthesis of A Water-Soluble Lipid I Analogue and A Convenient Assay for Translocase I. Analytical biochemistry, 461, 36-45.
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7

Evaluation of Endophytic Bacteria Inhibiting Fusarium

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A collection of 31 endophytic bacterial strains previously isolated from vetiver roots was used [24 (link)]. These strains were selected for their growth inhibition activity against F. graminearum in in vitro dual cultures and characterized taxonomically (Table 1). They were stored in a 15% glycerol solution at −80 °C. The 16S rDNA sequences of the 31 strains were deposited in GenBank, and the accession numbers are shown in Table 1.
The phytopathogenic fungal strains used in this study were F. graminearum, F. oxysporum, and F. culmorum. The F. graminearum strain was kindly provided by IFBM (Institut Français de la Brasserie et de la Malterie), while the other 2 strains were purchased from BCCM/MUCL: F. oxysporum f. sp. radicis-lycopersici Jarvis & Shoemaker (MUCL 039790) and F. culmorum (W.G. smith) Saccardo (MUCL 042823). The fungal strains were sub-cultured on potato dextrose agar (PDA) (OXOID) plates at 28 °C for F. graminearum and 25 °C for F. oxysporum and F. culmorum.
One strain of Escherichia coli K12 (ATCC, Manassas, MA, USA) and one strain of Saccharomyces cerevisiae DSM1333 (DSMZ, Braunschweig, Germany) were also used. Both were stored at −80 °C.
Munakata Y., Heuson E., Daboudet T., Deracinois B., Duban M., Hehn A., Coutte F, & Slezack-Deschaumes S. (2022). Screening of Antimicrobial Activities and Lipopeptide Production of Endophytic Bacteria Isolated from Vetiver Roots. Microorganisms, 10(2), 209.
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8

Bacterial Strain Preparation Protocol

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Mycobacterium tuberculosis (H37Rv) was obtained through BEI Resources, NIAID/NIH. Mycobacterium smegmatis (ATCC 607), and Escherichia coli K-12 (ATCC 29425) were obtained from ATCC. E. coli B21 wecA strain was obtained from Dr. McNeil (Colorado State University). A single colony of each bacterial strain was obtained on a Difco Middlebrook 7H10 nutrient agar enriched with 10% oleic acid, albumin, dextrose and catalase (OADC for M. tuberculosis), albumin, dextrose and catalase (ADC for M. smegmatis), and on Tryptic Soy agar (for E. coli). Seed cultures were obtained in Middlebrook 7H9 broth enriched with OADC (for M. tuberculosis), ADC (for M. smegmatis), and in Tryptic Soy broth (for E. coli), respectively. Each strain was grown to mid-log phase.
Mitachi K., Siricilla S., Yang D., Kong Y., Skorupinska-Tudek K., Swiezewska E., Franzblau S.G, & Kurosu M. (2016). Fluorescence-based Assay for Polyprenyl phosphate-GlcNAc-1-phosphate transferase (WecA) and Identification of novel antimycobacterial WecA inhibitors. Analytical biochemistry, 512, 78-90.
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9

Culturing Escherichia coli and Lactobacillus gasseri

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Escherichia coli K12 (American Type Culture Collection, VA), was grown in Luria-Bertani (LB) broth (BD Difco, MD) at 37 °C with shaking at 200 rpm. Lactobacillus gasseri ADH (generous gift from the Klaenhammer Lab, North Carolina State University)51 (link), was grown in De Man, Rogosa, Sharpe (MRS) broth (Thermo Fisher Scientific, NJ) at 37 °C.
Chen K.L., Liu X., Zhao Y.C., Hieronymi K., Rossi G., Auvil L.S., Welge M., Bushell C., Smith R.L., Carlson K.E., Kim S.H., Katzenellenbogen J.A., Miller M.J, & Madak-Erdogan Z. (2018). Long-Term Administration of Conjugated Estrogen and Bazedoxifene Decreased Murine Fecal β-Glucuronidase Activity Without Impacting Overall Microbiome Community. Scientific Reports, 8, 8166.
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10

Cultivation and Characterization of Diverse Enterobacteriaceae

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Escherichia coli K-12 (ATCC10798) was purchased from the American Type Culture Collection (ATCC; Virginia, USA). Citrobacter werkmanii (NBRC105721), Edwardsiella ictaluri (NBRC105724), Enterobacter hormaechei (NBRC105718), Erwinia persicina (NBRC102418), Pantoea agglomerans (NBRC102470), Pectobacterium carotovorum (NBRC14082), Proteus hauseri (NBRC3851), Serratia grimesii (NBRC13537), Trabulsiella guamensis (NBRC103172), and Yersenia bercovieri (NBRC105717) were purchased from the NITE Biological Resource Center (NBRC; Chiba, Japan). All bacterial strains were selected and handled based on the biosafety guidelines provided by our institution. E. coli was cultured at 37°C, E. persicina at 25°C while all other strains were cultured at 30°C with agitation. Luria Bertani (LB; 10 g/L Tryptone, 5 g/L yeast extract, 10 g/L NaCl, pH 7.0) broth was used as the culture medium. All experiments were performed with cells revived from cryostorage cultivated overnight for 16–20 h. Cells with an average OD600 of 1.0 measured with a Mini photo 518R (Taitec Inc., Tokyo, Japan) were used for subsequent experiments.
Toyohara D., Yokoi Y., Inoue G., Muraoka T, & Mori T. (2019). Abiotic Factors Promote Cell Penetrating Peptide Permeability in Enterobacteriaceae Models. Frontiers in Microbiology, 10, 2534.
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