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Streptococcus suis truncated protein Sao and application thereof

A technology for Streptococcus suis and truncated proteins, which can be applied in applications, antibacterial drugs, and veterinary vaccines. It can solve the problems of difficult purification of full-length Sao protein, obstacles to the development of Sao protein vaccines, and poor protein stability. It is not easy to achieve Degradation, stable properties, good stability

Active Publication Date: 2017-08-01
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The full-length Sao-M protein is 110KD. Wang Jing and others found that the purification of the full-length sao protein is difficult, the protein stability is poor, and the degradation rate is slow. These factors hinder the further development of Sao protein vaccines

Method used

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  • Streptococcus suis truncated protein Sao and application thereof
  • Streptococcus suis truncated protein Sao and application thereof
  • Streptococcus suis truncated protein Sao and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Synthesis of Streptococcus suis truncated protein Sao and vaccine preparation:

[0031] 1.1 Synthesis of truncated protein Sao

[0032] The sequence of the truncated protein Sao is shown in SEQ ID NO.2, which was synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. The truncated protein Sao polypeptide after synthesis has a purity of 99%, 5 mg each, and is in the form of white powder. It is stored in the dark at -20 degrees and is completely soluble in water. The protein was dissolved in PBS buffer solution, stored at -80°C for one month, and no protein degradation was found after taking it out.

[0033] Before use, wash with sterile PBS (NaCl 8.0g, KCl 0.2g, KH 2 PO 4 0.24g, Na 2 HPO 4 ×12H 2 O 3.628g, pH7.4) was diluted according to the specified concentration. Preheat in a 37-degree incubator before use.

[0034] 1.2 Preparation of vaccine

[0035] Dosage form: water-in-oil (W / O)

[0036] Adjuvant composition: white mineral oil (Marcol52), Tween 80 (C...

Embodiment 2

[0045] Application of Truncated Protein Vaccine Sao in Preparation of Streptococcus Suis Infection Vaccine

[0046] 1. Evaluate the immune protection effect of the vaccine on the mouse model

[0047] 1.1 Experimental scheme

[0048] Experimental animals: 4-week-old female BALB / c mice, purchased from Hubei Provincial Center for Disease Control and Prevention.

[0049] Challenge strain: Streptococcus suis type 2 SC19.

[0050] Experimental grouping: 20 female BALB / c mice weighing 16g were randomly divided into 2 groups: vaccine group I (Sao) and control group (adjuvant and PBS), 10 in each group. During the period after immunization, blood was collected by tail docking every week for the monitoring of serum-specific antibodies. Immunization method: subcutaneously immunize experimental mice with the vaccine prepared in Example 1 at a dose of 100 μl / only; booster immunization once 2 weeks later (immunization dose and site are the same as the first immunization).

[0051] 1.2 D...

Embodiment 3

[0074] Embodiment 3: Safety inspection

[0075] In order to test the safety impact of the prepared vaccine on piglets and pregnant sows, the safety test was carried out on 35-day-old piglets and 70-day-old sows.

[0076] 3.1 Experimental scheme:

[0077] 1) Inoculate 35-day-old healthy piglets with the prepared vaccine by point injection through the buttock muscle, inject 5 piglets in each batch of vaccine, and inoculate 2ml into each piglet, observe their clinical manifestations and measure their body temperature.

[0078] 2) The prepared vaccine was inoculated into healthy sows on the 70th day of gestation through hip muscle injection respectively, and each batch of vaccine was injected into 5 pigs, and each pig was inoculated with 2ml, and the clinical manifestations and litter conditions were observed.

[0079] 2. Test results

[0080] 3.2.1 The safety of the subunit vaccine of the present invention to the vaccination of weaned piglets

[0081] After the prepared vaccin...

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Abstract

The invention discloses streptococcus suis truncated protein Sao and application thereof. Amino acid sequence of the streptococcus suis truncated protein Sao is as shown in the SEQ ID NO.2. The protein stability is better than stability of full-length protein, and the protein of the invention is not easy to degrade. After the Sao protein is used for preparation of a vaccine, the vaccine can effectively protect piglets. By the use of large dose of virulent strain SC19 for counteracting toxic substances, the protecting force still reaches 60%. The Sao protein is safe and nontoxic. After immunization of piglets and pregnant sows, body temperature, breathe, appetite and metal state are all normal during the whole observation period, farrowing of pregnant sows is normal, and there is no abortion, fetal death and the like. The Sao protein can be combined with other proteins to prepare multi-vaccine and is suitable for mass production.

Description

technical field [0001] The invention relates to the field of livestock genetic engineering vaccines, in particular to a truncated protein Sao of Streptococcus suis and its application. [0002] technical background [0003] Streptococcus suis (S.suis) is the main pathogen that causes streptococcus suis. According to the capsular polysaccharide (CPS) on the surface of the bacteria, S.suis can be divided into 33 serotypes, namely type 1 / 2, 1 to 31 and 33 types. Among them, Streptococcus suis type 2 (SS2) is the serotype with the strongest virulence, wide prevalence and highest frequency of clinical isolation. SS2 is an important zoonotic pathogen that can cause porcine meningitis, endocarditis, sepsis, arthritis, serositis, pneumonia, etc., often causing sudden death of weaned piglets or fattening pigs; humans infected with this bacterium, It can lead to meningitis, which can lead to permanent deafness, sepsis, endocarditis, and even death. Suspected cases of Streptococcus s...

Claims

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Application Information

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IPC IPC(8): C07K14/315C12N15/31A61K39/09A61K39/116A61K39/02A61P31/04
CPCA61K39/0208A61K39/092A61K2039/552A61K2039/55511A61K2039/70C07K14/315
Inventor 金梅林李敬涛王雅康超吴超孙小美朱伟峰
Owner HUAZHONG AGRI UNIV
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