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Streptococcus mutans, application of streptococcus mutans and microbial preparation

A technology for Streptococcus mutans and microbial preparations, which can be applied to microorganisms, microorganisms, microorganism-based methods, etc., and can solve problems such as reports on the application of Streptococcus mutans antisense vicR gene mutants.

Active Publication Date: 2018-06-08
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] So far, there have been no reports on the construction of the antisense vicR gene mutant strain of Streptococcus mutans and its application in the prevention and treatment of dental caries

Method used

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  • Streptococcus mutans, application of streptococcus mutans and microbial preparation
  • Streptococcus mutans, application of streptococcus mutans and microbial preparation
  • Streptococcus mutans, application of streptococcus mutans and microbial preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] 1 Construction and identification of the antisense vicR gene mutant strain of Streptococcus mutans, the operation process is as follows figure 1 shown.

[0057] 1.1 Culture of strains

[0058] Streptococcus mutans UA159 (ie: Streptococcus mutans UA159, referred to as "S.mutans UA159") was provided by the State Key Laboratory of Oral Diseases Research of Sichuan University in Chengdu, Sichuan Province, China, and S.mutansUA159 was inoculated into bovine brain heart infusion (BHI) medium (Sigma-Aldrich Corp, Saint Louis, MO, USA), the bacteria were routinely subcultured overnight in a DY-2 anaerobic incubator (37°C, 5% CO 2 , 10%H 2 , 85%N 2 ; Zhejiang, China) for use.

[0059] 1.2 Construction of vicR antisense RNA recombinant expression vector

[0060] Through the NCBI database, the antisense RNA sequence (SEQ ID NO.1) was obtained, and the promoter sequence (SEQ ID NO. 7-358 in 3). Synthetic promoter + vicR gene antisense strand fragment nucleotide sequence (thi...

Embodiment 2

[0071] Detection of Growth Velocity of Streptococcus Mutans HT1171113

[0072] method:

[0073] Take the bacterial suspension of Streptococcus mutans UA159 and Streptococcus mutans HT1171113 in the mid-logarithmic growth period, and inoculate it in a 12-well plate with 2 mL of fresh BHI liquid medium (the final concentration of bacteria is 5×10 5 CFU / mL), at 37°C, 5% CO 2 Constant temperature cultivation in the incubator. At the beginning of the culture and at intervals of 1 hour, a well plate was taken out to measure the absorbance value at a wavelength of 600 nm, and the bacterial growth was monitored for 24 hours. Take the time point as the abscissa value, OD 600 The value is the ordinate value to draw the growth curve of the bacteria.

[0074] The result is as Figure 4 As shown, the time for Streptococcus mutans HT1171113 to reach the logarithmic growth phase was delayed compared with that of Streptococcus mutans UA159, indicating that the bacterial growth rate of St...

Embodiment 3

[0076] Detection of vicR antisense RNA expression content of Streptococcus mutans HT1171113

[0077]Total RNA extraction method: Streptococcus mutans UA159 and HT1171113 strains were inoculated in 5 mL of BHI and BHI (supplemented with 10 μg / mL spectinomycin) respectively, after overnight recovery, diluted in 10 mL of freshly prepared BHI containing 1% sucrose in a ratio of 1:20 After centrifugation, the bacterial samples were collected and resuspended in PBS for later use: ① Place on ice, operate intermittently for 60 s, and use FASTPREP Beater (MP Biomedicals, Irvine, CA) to break the cell wall thickness; Extract RNA. ②Layering: Add 1mL Trizol to the sample, pipette and mix repeatedly, then place it at room temperature for 5min to fully lyse the sample. Then add 200 μL of chloroform, mix vigorously up and down by hand for 15 sec, and then place at room temperature for 5 min to allow natural phase separation. ③ RNA precipitation: After centrifugation at 12000g at 4°C for 15...

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Abstract

The invention discloses streptococcus mutans, an application of the streptococcus mutans and a microbial preparation and relates to the field of prevention and treatment of caries. The streptococcus mutans is collected in China Center for Type Culture Collection (CCTCC) on number November 13, 2017, and a collection number of the streptococcus mutans is CCTCC NO: M2017746. The streptococcus mutanshas the characteristics of cariogenic relevant capability reduction such as decreased vicR gene expression and ViCR protein expression, a long chain shape, a decreased bacterium growth rate, decreasedVicR protein expression quantity, reduced biological membrane formation capability, reduced bacterial adherence capability and a reduced extracellular polysaccharide content.

Description

technical field [0001] The invention relates to the field of caries prevention and treatment, in particular to a streptococcus mutans and its application and microbial preparation. Background technique [0002] Caries is a chronic progressive infectious disease that occurs in the hard tissue of teeth, and its occurrence is closely related to the deposition of dental plaque biofilm. According to the results of the third oral epidemiological survey in my country, the dental caries rate of 5-year-old children and 35-44-year-old middle-aged people reached 66% and 88.1%, respectively, and the 65-74-year-old population was as high as 98%. The treatment rate is less than 10%. [0003] At present, more studies have pointed out that improper treatment of dental caries not only causes a series of oral problems, such as severe pain, loss of masticatory function, and affects facial appearance, but also is related to systemic diseases, such as digestive system diseases and cardiovascular...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N15/74C12N15/113A61K35/744C12R1/46
CPCA61K2035/115C12N1/20C12N15/113C12N15/746C12N2310/11C12N1/205C12R2001/46
Inventor 雷蕾杨英明毛梦莹陈虹张斌胡涛邓雅兰
Owner SICHUAN UNIV
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