Arnebia root endophyte as well as screening method, culture method and application thereof
A screening method, the technology of comfrey root, applied in the field of microorganisms, can solve the problems of unclear source of anti-colon cancer efficacy of comfrey root, unfavorable production of anti-colon cancer drugs, long planting time period and other problems, and achieve obvious anti-colon cancer effect, Potential medicinal value, good economic benefits
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0034] Example 1 Screening method
[0035] This embodiment provides a screening method for endophytes of comfrey roots: wash the comfrey roots with running tap water, use filter paper to absorb or dry the water on the comfrey roots, soak them in 75% ethanol for 3 minutes, and then soak them in 75% ethanol for 3 minutes. Soak the bacteria water three times for 3 minutes each time, then use 5% sodium hypochlorite solution to soak for 3 minutes, and finally use sterile water to wash 3~4 times to thoroughly clean the disinfectant. Aseptic scissors cut the comfrey root into thin sections of about 1 cm, and then placed it in a sterilized mortar for grinding and pulverizing, adding 1 ml of sterile water and mixing, and then moved to a shaking tube containing 5 ml of LB liquid medium. Then put it in a shaker for shaking culture, set the shaker temperature to 37°C and the rotation speed to 180rpm, incubate for 2~6h, take it out, let it stand for 5~10min, take 100μl of the upper liquid,...
Embodiment 2
[0036]Example 2 Identification of single colony species
[0037] Get the bacterial classification that has been preserved in the -80 ℃ refrigerator in the embodiment 1, activate the bacterial classification in the 100ml LB liquid medium according to the inoculation amount of 1% by volume, place the shaking culture in the shaking table, and set the temperature of the shaking table to be 37 ℃. ℃, the rotation speed is 225rpm, culture for 16h, after culture, put the bacterial liquid in a centrifuge tube, centrifuge in a refrigerated centrifuge, collect the bacterial species, use a genome extraction kit to extract the total genome DNA of the strain, and use the 16SrDNA universal primer for bacteria, Using the extracted genome as a template, PCR amplification was performed by denaturing at 95°C for 1 min, annealing at 50-60°C for 30s, and extending at 72°C for 2-5 min. The cycle was repeated 35 times. After PCR amplification, 1% agarose gel electrophoresis was performed. Electropho...
Embodiment 3
[0042] Example 3 The method of expanding the culture of strains
[0043] Get the comfrey root endophyte bacterial classification that has been preserved in -80 ℃ of refrigerators in embodiment 1, activate the bacterial classification in 100ml LB liquid medium according to the inoculation amount of 1% by volume, place in the shaker and carry out expanded culture, Set the shaker temperature to 37°C, the rotation speed to 225rpm, and cultivate for 12~24h, then place the fermentation broth in a centrifuge tube and use a refrigerated centrifuge for centrifugation. 5~12min, then remove the supernatant, collect the strain, then add 30ml of sterile PBS liquid, and then place it in a refrigerated centrifuge for centrifugation, remove the supernatant, and wash and remove the residual fermentation broth to obtain comfrey root endophytes. bacterial strains.
[0044] In order to identify the anti-colon cancer activity of comfrey root endophyte, the present invention prepares the comfrey r...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com