Application of chelerythrine in inhibition of colistin drug resistance gene transfer
A technology of chelerythrine and drug-resistant genes, applied in the field of biomedicine, can solve problems such as limiting the application of colistin, achieve wide application value, and inhibit the spread of the effect
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[0029] The preparation method of the culture medium involved in the following examples is as follows:
[0030] BHI broth medium: add 18.5 g of LB broth medium (Beijing Land Bridge Technology Co., Ltd.) to an appropriate amount of distilled water, then dilute to 500 mL with distilled water, sterilize by autoclaving at 121°C for 20 min, and cool.
[0031] BHI agar medium: add 25 g of BHI agar medium (Beijing Luqiao Technology Co., Ltd.) to an appropriate amount of distilled water, then dilute to 500 mL with distilled water, sterilize by autoclaving at 121 °C for 20 min, pour the plate, and cool.
Embodiment 1
[0032] Example 1. Identification of Escherichia coli ZJ807, Escherichia coli ZJ28 and Escherichia coli J53
[0033] Escherichia coli ZJ807 was described in the following documents: Prevalence, risk factors, outcomes, andmolecular epidemiology of mcr-1-positive Enterobacteriaceae in patients and healthy adults from China: an epidemiological and clinical study), and the genome sequence was uploaded to NCBI, BioSample Accession Number: SAMN05437814.
[0034] Escherichia coli ZJ28 was described in the following documents: Prevalence, risk factors, outcomes, andmolecular epidemiology of mcr-1-positive Enterobacteriaceae in patients and healthy adults from China: an epidemiological and clinical study, and the genome sequence was uploaded to NCBI, BioSample Accession Number: SAMN05437813 .
[0035] Escherichia coli J53 is described in: Genome sequence of Escherichia coli J53, a reference strain for genetic studies. GenBank accession number AICK00000000.
[0036] It was identified t...
Embodiment 2
[0037] Example 2. Application of chelerythrine in inhibiting the transfer of mcr-1 positive plasmids
[0038] Using Escherichia coli ZJ28 or Escherichia coli ZJ807 as the donor strain and Escherichia coli J53 as the recipient strain, the effect of chelerythrine on the transfer of mcr-1 positive plasmids between Escherichia coli was evaluated by conjugation transfer method.
[0039] 1. Preparation of polymyxin E stock solution, chelerythrine stock solution and sodium azide stock solution
[0040] (1) get polymyxin E, dilute with deionized water, obtain the polymyxin E solution with a concentration of 2560 μg / mL; then use the polymyxin E solution with a sterile filter membrane (Pall company in the United States, the pore size is 0.22 μm) to obtain a stock solution of polymyxin E.
[0041] (2) Take chelerythrine and dilute it with deionized water to obtain a chelerythrine solution with a concentration of 1000 μg / mL; then filter the chelerythrine solution with a sterile filter to...
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