Streptomyces pratensis and method for producing pseudouridine and 1-methyl-pseudouridine by fermenting streptomyces pratensis
A technology of Streptomyces platetts and pseudouridine, applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., which can solve the problems of insufficient preparation methods
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Embodiment 1
[0032] Example 1: Strain source
[0033] Streptomyces platensis DHE020 was isolated from the soil of Taihe Mountain in Danjiangkou City, Hubei Province, China.
[0034] Cross-sampling the soil in the Taihe Mountain area, randomly select 5 sampling points, take 10g of soil sample from each point, put it into a conical flask, mix it evenly, take 10g of the sample, and add it to a cone filled with 90mL of sterile water In a shaped bottle (there is a magnetic stirrer in the bottle), vortex stirring for 30 minutes to make it fully mixed to form a suspension, that is, 10 -1 Bacterial suspension. Dilute the above suspension with sterile water according to the volume ratio of 1:9 to 10 by the dilution coating plate method. -2, 10 -3 , 10 -4 , 10 -5 Concentration, take 0.1 mL of bacterial suspension of different dilution multiples, spread it on ISP2 medium plate, gently spread on the surface of the medium with a sterile coating rod, let stand for 30 minutes at room temperature and...
Embodiment 2
[0037] Example 2: Morphology, culture characteristics, physiological and biochemical characteristics of Streptomyces platensis DHE020.
[0038] Refer to "Streptomyces Identification Manual", "Classification and Identification of Actinomycetes", "Common Bacteria System Identification Manual" and other books to carry out experiments: the color judgment is compared with the color in the RAL K7 color card.
[0039] 1. Morphological characteristics of the strain: The strain DHE020 was inoculated in ISP2 medium for insert culture, and after culturing at 28°C for 3-5 days, the cover glass was taken into the slide and observed under an optical microscope at 400× magnification. The results are shown in figure 1 .
[0040] 2. Culture characteristics of strains: After strain DHE020 was cultured on ISP2 medium at 28°C for 7-10 days, the colony was oval in shape, with radial stripes, raised on the surface, the diameter of the colony was about 8-20mm, and there were grooves on the surface ...
Embodiment 3
[0065] Example 3 Identification of bacterial species
[0066] 1. 16S rDNA sequence analysis of Streptomyces platter DHE020
[0067] Carry out experiments with reference to the relevant content in the book "Molecular Cloning Experiment Guide". The mycelia were collected, and then total DNA was extracted with an actinomycete DNA extraction kit. The universal primer 27F (2 7F: 5'-AG AG T TT G AT C C T G G C TC A G -3') / 1495R (1495R 5'-CTACGGCTACCTTGTTACGA-3') was used to amplify the 16S rDNA sequence, and the PCR products were detected by 0.8% agarose Gel electrophoresis, PCR products were purified and recovered using the SanPrep column-type PCR purification product kit, and the purified PCR products were directly sent to Nanjing GenScript Biotechnology Co., Ltd. for sequence determination.
[0068] After proofreading, the 16S rDNA sequence of strain DHE020 was compared with the sequences of related species and genera in GenBank database by BLAST comparison of homologous sequen...
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