Light-emitting reporting system and application thereof to antibiotic discovering and screening
A reporting system and antibiotic technology, applied in the biological field, can solve problems such as antibiotic discovery and screening that have not yet been achieved, and achieve the effects of increasing probability, speeding up discovery, and huge social and economic benefits
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Embodiment 1
[0025] Experimental strains and plasmids:
[0026] Escherichia coli DH5α: used for competent preparation, obtained from China General Microorganism Culture Collection Center (CGMCC1.12873).
[0027] Escherichia coli DH10B: used for competent preparation, obtained from ThermoFisher Scientific (Cat. No. 18297010)
[0028] Staphylococcus aureus RN4220: used for competent preparation and luminescence detection, acquired from BEI Resources.
[0029] pMD19-T: Purchased from Dalian Bao Biological Company (TaKaRa)
[0030] pAmilux: from the University of British Columbia, Canada (Document: Mesak LR, Yim G, Davies J, Improved lux reporters for use in Staphylococcus aureus, Plasmid, 2009, 61(3): 182-187.)
[0031] The medium and its preparation required for the experiment:
[0032] LB medium: For each liter of medium, tryptone (10g), sodium chloride (10g), and yeast extract (5g) should be added to 950ml of deionized water, and stirred with a magnetic stirrer until dissolved. Add 5 m...
Embodiment 2
[0146] Example 2 The response of the luminescent reporter system to antibiotics
[0147] Experimental strains: The strains used in the experiment are four lux reporter strains of Staphylococcus aureus: S1, S2, S3, S4, and the strain S0 containing pAmilux empty plasmid.
[0148] Media required for experiment and its preparation
[0149] 1. NYE agar medium
[0150] 2. 0.7% agar: Weigh 0.7g of agar with a balance and add it to the Erlenmeyer flask, then add 100ml of deionized water, and sterilize under high pressure steam at 121°C for 20 minutes for later use.
[0151] Antibiotics required for experiments and their preparation
[0152] 1. Daptomycin (DAP) solution: Dissolve 30 mg of daptomycin in 1 ml of ultrapure water to make the final concentration 30 mg / ml, and filter through a 0.22 μm membrane to sterilize;
[0153] 2. Polymyxin B (PMB) solution: Dissolve 100mg of polymyxin B in 1ml of ultrapure water to a final concentration of 100mg / ml, and filter through a 0.22μm membr...
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