Bacillus cereus phage and application thereof
A technology of Bacillus cereus and bacteriophage, applied in the direction of phage, virus/phage, application, etc., can solve the problems of limited phage and inability to kill Bacillus cereus, and achieve good thermal stability
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Embodiment 1
[0030] Example 1. Preservation information of bacteriophage
[0031]The present invention provides a phage, which is named as Bacillus cereus bacteriophage DLc1, and Bacillus cereus bacteriophage DLc1 has been preserved in Guangdong Microbiological Culture Collection on September 18, 2020 Center, address: Guangdong Institute of Microbiology, No. 100, Xianlie Middle Road, Guangzhou, China, and the deposit number is GDMCC NO: 61196-B1.
Embodiment 2
[0032] Example 2, Isolation, Enrichment and Purification of Phage
[0033] The separation and purification of the phage of the present invention comprises the following steps:
[0034] 1. Preparation of bacterial suspension: Bacillus cereus 1582-3B carrying multiple virulence genes and multidrug resistance was used as a sensitive indicator strain, and Bacillus cereus 1582-3B was a pasteurized milk isolate. Freeze in a glycerol tube and activate by streaking on a plate to make a bacterial suspension.
[0035] 2. Separation of bacteriophage: Collect sewage water samples collected from Huangsha Aquatic Products Market in Liwan District, Guangzhou City, Guangdong Province. First, the sewage water samples are centrifuged at 8,000×g for 10 minutes to remove large particles such as sediment, and then filtered through a 0.45 μm pore size filter membrane. Most of the environmental bacteria in the water sample; then add magnesium sulfate to a final concentration of 50 mM, let it stand ...
Embodiment 3
[0039] Embodiment 3, preparation of high-titer phage stock solution
[0040] Pick out the single phage plaque that has been purified to a uniform shape in Example 2, resuspend it in 3 mL of TSB broth medium containing 1 mM calcium chloride, and inoculate it with 1% activated Bacillus cereus 1582-3B strain, at 37°C Cultivate under shaking for 3 hours, centrifuge at 10,000×g for 1 minute, and then filter to obtain the supernatant; take another 3 mL of TSB broth medium containing 1 mM calcium chloride, and inoculate 1% Bacillus cereus 1582-3B activated strain 37 Cultivate with shaking at ℃ for 1 hour, then add 100 μL of the supernatant obtained above, continue to shake and co-culture at 37°C for 6 hours, centrifuge at 10,000 g for 1 minute, and filter to obtain the supernatant after preliminary amplification; take a freshly prepared 50 mL containing 1 mM chloride Calcium TSB broth medium, inoculated with 1% activated Bacillus cereus 1582-3B strain, cultured with shaking at 37°C f...
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