Micromolecular substance for improving sensitivity of bacteria to antibiotics
A technique for antibiotics, susceptibility, used in the field of medicine
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Embodiment 1
[0037] Screening of antibiotic resistant strains of Escherichia coli K12 BW25113
[0038] Escherichia coli ( Escherichia coli , E. coli )) is the most common type of bacteria in the intestinal tract of humans and animals, mainly in the large intestine, accounting for about 1% of intestinal bacteria. Detect the starting bacteria of Escherichia coli ( Escherichia coli K12 BW25113) minimum inhibitory concentration (MIC) for 10 antibiotics. Will 10 5 The starting bacteria of colony forming unit / ml were cultured in 10 kinds of LB liquid medium containing 1 / 2 of the minimum inhibitory concentration of antibiotics at 37°C and successively passaged 10 times, and the minimum inhibitory concentration of the obtained single clone was determined. The results showed that the minimum inhibitory concentrations of the 10 selected bacteria against the 10 antibiotics were 64 times the minimum inhibitory concentrations of the respective starting bacteria (see Table 1), indicating that 10 types...
Embodiment 2
[0043] Glutamine is a key biological target substance of ampicillin-resistant Escherichia coli
[0044] 1. Preparation of GC-MS samples of ampicillin-resistant Escherichia coli
[0045] 1) Preparation of bacterial samples: pick the starting bacteria of Escherichia coli from the solid LB plate ( Escherichia coli K12 BW25113) and a single colony of ampicillin-resistant bacteria (see Example 1 for the specific method of culturing resistant bacteria), inoculate in 5 ml of LB medium, culture at 37°C and 200 rpm for 16 hours; press 1:100 (v / v) Inoculated in 100 ml of LB liquid medium, and cultivated at 37°C to an OD600 value of 1.0.
[0046] 2) Preparation of GC-MS samples: Take 10 ml of bacterial solution and quickly add 5 ml of frozen methanol to quench the sample to stop the metabolic process of cells. Collect the bacteria by centrifugation at 4°C and 8000rpm for 3 minutes, remove the water as much as possible, and store the bacteria in 2 ml of cold methanol (Sigma).
[0047] 3) Ext...
Embodiment 3
[0054] Glutamine down-regulation is widespread in various antibiotic-resistant bacteria of Escherichia coli
[0055] 1. Preparation of GC-MS samples of various antibiotic-resistant Escherichia coli bacteria
[0056] Cultivation of Escherichia coli starting bacteria ( Escherichia coli K12 BW25113) and its 10 resistant E. coli strains: GEN-R, AMP-R, BLFX-R, KAN-R, NA-R, CAZ-R, ER-R, CAP-R, SM-R and TET -R, refer to Example 2.1 for the sample preparation process
[0057] 2. The down-regulation of glutamine in various antibiotic-resistant bacteria of Escherichia coli is a common phenomenon
[0058] The prepared 10 kinds of drug-resistant bacteria samples were loaded on GC-MS to analyze metabolites. According to the retention time of glutamine in the GC-MS total ion flow chart, the peak area data of the glutamine peak and the internal standard peak of each drug-resistant bacteria are obtained, and the relative peak area (ratio of the internal standard peak) is used to represent the m...
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