Drug-resistant pathogen infection resistant polypeptide and uses thereof
An antibacterial polypeptide and drug resistance technology, applied in the field of biomedicine, can solve the problems of accelerating the evolution of pathogenic bacteria and increasing the rate of bacterial drug resistance, and achieve the effect of high antibacterial activity
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Embodiment 1
[0027] Synthesis and structure confirmation of peptides:
[0028] The polypeptide of the present invention is artificially synthesized by conventional solid-phase synthesis method. Polypeptide Cbf-K of the present invention 16 and Cbf-A 7 A 13 , its purity determined by HPLC is greater than 99%, and its molecular weight determined by mass spectrometry is 3637.63 and 3496.37 respectively, and its molecular weight is consistent with the theoretical value of 3638.1 and 3496.8.
Embodiment 2
[0030] Polypeptide Cbf-K of the present invention 16and Cbf-A 7 A 13 In vitro antibacterial effect on recombinant E.coli NDM-1:
[0031] (1) Accurately weigh the polypeptide Cbf-K 16 and Cbf-A 7 A 13 Prepare a drug solution with a concentration of 1280 μg / ml, filter it aseptically with a 0.22 μm membrane, aliquot it, and store it at -70°C for later use.
[0032] (2) Inoculate recombinant E.coli NDM-1 on the slant of nutrient agar, incubate at 37°C for 18h, then pick a little and inoculate it into 2ml nutrient broth medium, incubate at 37°C for 8h, dilute with sterile MH broth into 10 5 Bacterial suspension with about CFU / ml was continuously induced with IPTG.
[0033] (3) Take a sterile 96-well plate, add 40 μl of drug stock solution and 60 μl of MH broth medium to the first column, add 50 μl of MH broth medium to each well of the remaining wells, and dilute the drug in turn to 50 μl containing drug in each row Solutions with concentrations of 512, 256, 128, 64, 32, 16...
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