Bacillus marinus and application thereof
The technology of marine bacillus and bacillus, which is applied in the field of biomedicine, can solve the problems of reducing the quality of life of cancer patients, specifically targeting tumor cell division obstacles, etc., and achieve the effect of malignant tumor prevention and treatment
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Embodiment 1
[0036] Embodiment 1, the acquisition of marine bacillus:
[0037] In March 2016, followed the "Science" to collect sea mud samples in the seamount area of the western Pacific Ocean (139°3802'E, 11°44162'N). The collected samples were serially diluted with sterile distilled water, and 10 -6 ,10 -5 ,10 -4 0.2 ml each of the diluted solution was spread on the solid medium, and cultured at 28 degrees for 48 hours. The colonies were transferred to fresh solid medium (5g tryptone, 1g yeast extract in 1L sterile seawater, and 15 agars were added) by the tetrad streak method, and transferred several times until the pure culture of the strain was obtained. This strain is stored in the General Microbiology Center of China Microbiological Culture Collection Management Committee, address: No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing (Institute of Microbiology, Chinese Academy of Sciences), and the preservation date is December 2017. , deposit number: CGMCC NO.14928. ...
Embodiment 2
[0042] Preparation of Exopolysaccharide from Bacillus sp.11
[0043] Pick a single colony and inoculate it into a test tube containing 5ml of liquid 2216E medium, and culture it with shaking at 28°C and 160rpm for 24h to obtain a seed fermentation broth. The seed fermentation broth of the Bacillus was inoculated into 500 ml of 2216e medium with 1% sucrose at a ratio of 0.1%, and cultured at 28° C. and 160 rpm for 48 hours with shaking. Centrifuge (8000rpm, 20min) to remove the bacteria, then add 3 times the volume of supernatant 95% ethanol solution to precipitate overnight at 4°C, then centrifuge (8000rpm, 20min, 4°C) to take the precipitate, and add water to dissolve all the precipitate. After dissolving, the protein was removed by Sevage reagent three times, and then the solution was put into a dialysis bag of 8000-14000Da for dialysis. The dialysate was filtered through a 0.22μm filter membrane, condensed with an anion column, and eluted with 20mM Tris and 2M NaCl. The e...
Embodiment 3
[0046] Effects of EPS11 administered in vitro on the proliferation activity of lung cancer A549 cells
[0047] experimental method
[0048] Take the A549 cells in the logarithmic growth phase, digest the cell solution in the logarithmic growth phase into a single cell suspension with trypsin, count the cells, and divide the cells according to the ratio of 6×10 3 The density of cells / well was seeded in 96-well cell culture plate, and placed in 5% CO 2 , 37°C cell culture incubator; the next day, add EPS11 with final concentrations of 0, 0.01, 0.02, 0.04, 0.10, 0.20, 0.50, 1.00 and 2.00mg / ml to the 96-well plate, and set the concentration of each group 3 duplicate wells were put into the incubator to continue culturing for 24h and 48h respectively. After being treated with EPS11, 30μl of MTT solution (concentration: 5mg / ml) was added to each well, placed in the incubator and then cultured for 4h, and then each well was Then add 100 μl of triple solution, put it into the incuba...
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