Lactobacillus salivarius strain and application thereof
A technology of Lactobacillus salivarius and strains, applied in the field of microorganisms, can solve the problems of natural lactobacillus intolerance to high temperature and the death of lactobacillus
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Embodiment 1
[0020] The isolation of embodiment 1 bacterial strain
[0021] experiment material:
[0022] Test sample: In the middle of the Tianshan Mountains in Xinjiang, China, the surface humus layer with inaccessible and dense vegetation
[0023] Preparation of medium: MRS liquid medium: peptone 10g, beef extract 10g, yeast extract 5g, diammonium hydrogen citrate 2g, glucose 20g, Tween-801ml, sodium acetate 5g, dipotassium hydrogen phosphate 2g, magnesium sulfate 0.58g , manganese sulfate 0.25g, distilled water 1000ml, pH value 6.2-6.4, sterilized at 121°C for 15min;
[0024] MRS solid medium: MRS liquid medium is prepared by adding 1.5% agar powder and sterilizing at 121°C for 15 minutes.
[0025] experiment method:
[0026] The collected humus layer was directly inoculated into the MRS liquid selection medium for bacterial proliferation and enrichment, and then inoculated into the MRS solid selection medium, and the colonies with obvious characteristics were picked and separated b...
Embodiment 2
[0029] Biochemical identification and physicochemical characteristics of embodiment 2 bacterial strains
[0030] Test materials: Bovine bile salt reagent was purchased from Aoboxing Biotechnology Co., Ltd.
[0031] Biochemical identification of strains:
[0032]Use the API50CHL test strip identification system of French Mérieux company to identify the biochemical profile of the strain, and carry out the species identification of the strain according to the biochemical profile (API50CHL is an identification test strip system for Lactobacillus and related bacteria, which is composed of 49 fermentable A test strip composed of API50CH simple medium of carbohydrates. Each small tube of the test strip is inoculated with the suspension inoculated with the measured bacteria. When cultivating, due to the acid production of fermented carbohydrates, the pH drops, causing the indicator to change color. The result constitutes the strain Biochemical map, and used for identification or typi...
Embodiment 3
[0037] The 16srDNA species identification of embodiment 3 bacterial strains
[0038] 1 Test material:
[0039] The PCR kit was purchased from TaKaRa Biological Company; the gel recovery kit was purchased from Ai Sijin Biotechnology Co., Ltd.; the primers were synthesized by Boshi Bio Company, using the bacterial domain universal primer 27F as SEQ ID No. 2, and 1541R as SEQ ID No. 3.
[0040] Using the general primers 27F and 1541R in the bacterial domain, the extracted Lactobacillus genomic DNA was used as a template to amplify the 16S rDNA sequence with a length of 1468 bp. The PCR amplification system was 50 μL. PCR program: 95°C for 5min; 95°C for 45s, 50°C for 1min , 72°C 90s, 10 cycles; 95°C 45s, 58°C 1min, 72°C 90s, 15 cycles; 95°C 45s, 55°C 1min, 72°C 90s, 10 cycles; 72°C 10min, PCR amplification product cut After the gel was recovered, the fragment was cloned into the pMD18-T vector and sequenced by Invitrogen Company. The obtained sequence was more than 99% homologo...
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