Application of amentoflavone in preparation of staphylococcus aureus MgrA inhibitor
A kind of technology of biflavone and staphylococcus, which is applied in the direction of antibacterial drugs, medical preparations containing active ingredients, and pharmaceutical formulas, etc.
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Embodiment 1
[0016]Inhibition of MGRA of Pimple Sapulfone Transcriptional Factor MGRA
[0017]The PET28A-MGRA recombinant plasmid was converted to E. coli expression strain BL21 (DE3) by a thermosomycin-resistant screening, and then sequencing the Single colony by PCR identification from the plate. The sequencing strain was seeded into the LB medium, and at 37 ° C to OD600 = 0.6 to 0.8, IPTG was added to induce recombinant protein at 16 ° C for IPTG at 37 ° C, and the final concentration of 0.5 mmol / L was added to induce recombinant proteins. The bacterium that was collected overnight culture was collected, and the ultrasonic crushing was purified by nickel pillars and chromatographic techniques for purification of MGRA proteins with HIS labels. The protein was eluted using 20 mmol / L of imidazole buffer washing protein, and the protein was collected, and the protein was concentrated, and the protein concentration was determined by nucleic acid protein quantitator NanoDROP.
[0018]Inhibitory effec...
Embodiment 2
[0021]Effects of Pimple Doublelavone on the Growth of Plasticis USA300
[0022]In order to further determine the effect of the growth of Pine Splastic S.cocystis to the growth of the Plasticis USA 300, the overnight culture is cultured in a sterile BHI medium in a sterile BHI medium in a proportion of 1: 100, and different concentrations (0 μg / ml) , 64 μg / ml, 128 μg / ml, 256 μg / ml) of panfozine, 37 ° C oscillated culture for 24 h. Setting the MGRA lack of mutant strains and only the DMSO group was used as a control. The OD value at 600 nm was measured at different time points using the UV spectrometer, and the growth curve was drawn using the PRISM 7.0 software.
[0023]Resultsfigure 2 As shown, even if the dosage of the sputum double flavonoid is its IC5080 times, the growth of S. aureus is not significantly affected. The results showed that the activity of Pine S. flavonoids can inhibit the activity of a Staphylococcus aureus transcriptional regulatory factor MGRA without affecti...
Embodiment 3
[0025]Sapulatin inhibits the adhesion of Fibrinogen in Pimple Sapulfone
[0026]The activated Golden Staphylococcus USA 300 was inoculated in a BHI medium containing 3% NaCl and 0.5% glucose in a ratio of 1: 100, and different concentrations of panfosa double flavonoids (16 μg / ml, 32 μg / ml, 64 μg) were added. / ml, 128 μg / mL, 256 μg / mL) and a control group of unharacterized drugs. USA300 in 37 ° C constant temperature oscillation into OD600= 0.5, centrifugation collected bacteria and washed 3 times with PBS, followed by the bacterial herescent, and adjusted to OD600= 1.0. The 96-well plate was filled with a 96-well plate with 20 μg / ml fibrinogen at 4 ° C, and 5% BSA solution was added after 2 h after flushing with PBS. The resuspended bacteria was added in a packet according to the group, and the three pores were repeated each set. After 37 ° C for 1 h, 100 μl of 25% formaldehyde was added 20 min, and finally crystalline purple dye was decolorized with ethanol, and the OD val...
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