Streptococcus mutans rnc gene mutant strain and application thereof
A technology for Streptococcus mutans and strains, which is applied in bacteria, microorganism-based methods, and medical preparations containing active ingredients, etc., can solve problems such as reports on the application of Streptococcus mutans rnc gene mutant strains, etc., and reduce the occurrence of rate, reduction of caries, and low cariogenic potential
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Embodiment 1
[0036] Example 1: The acquisition and identification of "Streptococcus mutans HT120211CCTCC M2013211" of the present invention, see the procedure figure 1
[0037] 1. Construction and identification of "Streptococcus mutans HT120211CCTCC M2013211" of the present invention
[0038] 1.1 Cultivation of strains
[0039] Streptococcus mutans UA159 (ie: Streptococcus mutans UA159, referred to as "S.mutans UA159") obtained from the State Key Laboratory of Oral Disease Research in Chengdu, Sichuan Province, China was inoculated into bovine brain heart infusion (BHI) medium ( Sigma-Aldrich Corp, Saint Louis, MO, USA), the bacteria were routinely subcultured overnight and cultured in an DY-2 anaerobic incubator (37°C, 5% CO2, 10% H2, 85% N2; Zhejiang, China) for later use.
[0040] 1.2 Construction of Streptococcus mutans HT120211 mutant strain
[0041] Check the NCBI database to obtain the DNA sequence of the rnc gene and its upstream and downstream fragments, and design primers (rnc-P1, rnc-P2...
Embodiment 2
[0074] Example 2: Comparison of the cariogenic ability of "Streptococcus mutans HT120211CCTCC M2013211" of the present invention in a rat model of human caries and the male parent strain Streptococcus mutans UA159
[0075] 1. In the rat model of human dental caries, Streptococcus mutans UA159 and Streptococcus mutansHT120211CCTCC M2013211 were inoculated into 10ml BHI (supplemented with 1% sucrose) overnight, centrifuged, and the supernatant was discarded and resuspended in 10ml fresh BHI medium. Aspirate 100 μl of the bacterial suspension in 2ml BHI medium and continue to culture to an optical density value of 0.4 (OD600).
[0076] In the in vivo experiment, the animals were kept for 3 weeks after the successful colonization of the strains, CO 2 The rats were killed by asphyxiation method, the mandible specimens were taken, and sterilized normal saline was gently rinsed 3 times. The right mandible was placed in a sterile 15ml test tube containing 5ml RNA tissue sample protection s...
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