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Streptococcus mutans surface protein antigen (SpaP) and glucosyltransferase (GtfB) fused protein vaccine and preparation method thereof

A technology of glucosyltransferase and Streptococcus mutans, which is applied in the directions of transferase, bacterial antigen components, chemical instruments and methods to achieve good development and application prospects and good immunogenicity.

Active Publication Date: 2015-09-30
武汉第一口腔医院有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, preliminary results have been obtained in the research and development of the surface antigen (SpaP) and GtfB (GlU region) peptide vaccines of Streptococcus mutans. It is found that these two protein peptide vaccines can provide certain protection against dental caries, but how to further improve Its immune effect is a big challenge

Method used

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  • Streptococcus mutans surface protein antigen (SpaP) and glucosyltransferase (GtfB) fused protein vaccine and preparation method thereof
  • Streptococcus mutans surface protein antigen (SpaP) and glucosyltransferase (GtfB) fused protein vaccine and preparation method thereof
  • Streptococcus mutans surface protein antigen (SpaP) and glucosyltransferase (GtfB) fused protein vaccine and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0039] [Example 1] Construction of Spap and SpaP-(G4S)3-GtfB recombinant protein

[0040] 1. Design and synthesis of primers

[0041]The surface protein SpaP sequence (Genbank Gene ID: 102805) and the β-1,3-glucanase GtfB coding sequence (Gene ID: 1028336) from the genome sequence of Streptococcus mutans UA159 in NCBI:BLAST , design and synthesize three pairs of specific primers, the primers are synthesized by Shanghai Sangon Bioengineering Technology Co., Ltd., the sequences and numbers of the primers are as follows:

[0042] 1) GtfB gene PCR amplification primers

[0043] Upstream primer P1 (SEQ ID NO.2)

[0044] 5′-CGC GGATCC ATGGGCTATCAAGCCAAAG-3′

[0045] Downstream primer P2 (SEQ ID NO.3)

[0046] 5′-CCG CTCGAG AATCCGAACTCGTTTCCAG-3′

[0047] (The 5' end of the amplification product contains a BamHI site, and the 3' end contains an XhoI site)

[0048] 2) SpaP-(G4S)3 gene PCR amplification primers

[0049] Upstream primer P3 (SEQ ID NO.4)

[0050] 5′-CTA GCTAGC AT...

Embodiment 2

[0127] [Example 2] Detection of protein vaccine immune efficacy of SpaP and SpaP-(G4S)3-GtfB recombinant protein nasal cavity immunized mice

[0128] To study the immune effects of two anti-caries subunit vaccines in mice, and to verify and compare the immune efficacy of SpaP and SpaP-(G4S)3-GtfB proteins. The present invention uses BALB / c mice as animal models.

[0129] 1. Vaccine safety evaluation of SpaP and SpaP-(G4S)3-GtfB protein vaccines

[0130] Fifteen healthy female BALB / c mice aged 5-6 weeks were randomly divided into 3 groups, with 5 mice in each group. The mice in groups A and B were intramuscularly injected with SpaP and SpaP-(G4S)3-GtfB protein vaccines (dose 10 μg / 100 μl per mouse), mice in group C were intramuscularly injected with 100 μl sterile PBS as a negative control.

[0131] The mice were observed for 21 days after injection, and the weight change was measured. On days 5, 7, 9, 14 and 21, blood was collected from the tail and the number of white blood...

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Abstract

The invention relates to a streptococcus mutans surface protein antigen (SpaP) and glucosyltransferase (GtfB) fused protein vaccine and a preparation method and application thereof. The amino acid sequence of a fused protein is SEQ ID NO:1. SpaP genes and GtfB genes are connected according to a Lingker sequence to obtain the fused protein which is used for preparing a caries prevention protein vaccine. The genetic engineering sigmasubunit vaccine is good in immunogenicity and can efficiently cause an immune response of an inoculum.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a Streptococcus mutans cell surface protein antigen (SpaP) and glucosyltransferase (GtfB) fusion protein vaccine and a preparation method thereof. Background technique [0002] Dental caries, cardiovascular disease and cancer are called the three major human diseases. In China, 80% of children, 50% of adults, and 98% of the elderly suffer from it to varying degrees. To achieve effective prevention of dental caries, in addition to reducing carbohydrate intake, using fluoride to enhance tooth resistance to acid, and using sealants to reduce the colonization of pathogenic bacteria in tooth grooves, the use of antibacterial substances to reduce or eliminate cariogenic microorganisms is a must. The focus of scientists' research for decades, the research and development of dental caries vaccine is the top priority, but so far, there is still no vaccine that can be used clinica...

Claims

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Application Information

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IPC IPC(8): C07K19/00C07K14/315C12N9/10C12N15/70A61K39/09A61P1/02A61P31/04
Inventor 樊明文李宇红许庆安贾荣曹茜茜
Owner 武汉第一口腔医院有限责任公司
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