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A kind of streptococcus suis truncated protein sao and its application

A technology of Streptococcus suis and truncated protein, applied in application, antibacterial drugs, veterinary vaccines, etc., can solve the problems of difficult purification of full-length sao protein, hindering the research and development of Sao protein vaccine, poor protein stability, etc. It is not easy to achieve Degradation, stable properties and good stability

Active Publication Date: 2018-11-06
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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  • A kind of streptococcus suis truncated protein sao and its application
  • A kind of streptococcus suis truncated protein sao and its application
  • A kind of streptococcus suis truncated protein sao and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The synthesis and vaccine preparation of the truncated protein Sao of Streptococcus suis:

[0030] 1.1 Synthesis of truncated protein Sao

[0031] The sequence of the truncated protein Sao is shown in SEQ ID NO. 2, synthesized by Shenggong Bioengineering (Shanghai) Co., Ltd. After synthesis, the truncated protein Sao polypeptide has a purity of 99%, 5mg each, in white powder form, stored at -20 degrees in the dark, and is completely soluble in water. The protein was dissolved in a PBS buffer solution, stored at -80°C for one month and then taken out and no degradation of the protein was found.

[0032] Before use, use 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) Dilute according to the specified concentration. Preheat in a 37 degree incubator before use.

[0033] 1.2 Preparation of vaccine

[0034] Dosage form: water-in-oil type (W / O)

[0035] Adjuvant composition: white mineral oil (Marcol52), Tween 80 (CRILLET4), Span 80 (CRILL4).

[0...

Embodiment 2

[0046] Application of truncated protein vaccine Sao in preparation of vaccine against Streptococcus suis infection

[0047] 1. Evaluate the immune protection effect of the vaccine on a mouse model

[0048] 1.1 Experimental protocol

[0049] Experimental animals: 4-week-old female BALB / c mice, purchased from Hubei Provincial Center for Disease Control and Prevention.

[0050] Challenge strain: Streptococcus suis type 2 SC19.

[0051] Experimental grouping: 20 female BALB / c mice weighing 16g were randomly divided into 2 groups: vaccine group I (Sao), control group (adjuvant and PBS), 10 mice in each group. During the post-immunization period, blood was collected from the tail every week for monitoring of serum specific antibodies. Immunization method: The vaccine prepared in Example 1 was subcutaneously immunized with a dose of 100 μl / mouse; a booster immunization was performed 2 weeks later (the dose and location of the immunization were the same as the first immunization).

[0052] 1.2...

Embodiment 3

[0075] Example 3: Safety inspection

[0076] In order to examine the safety effects of the prepared vaccine on piglets and pregnant sows, safety inspections were carried out on 35-day-old piglets and 70-day-old sows.

[0077] 3.1 Experimental program:

[0078] 1) Inoculate 35-day-old healthy piglets through separate injections of the prepared vaccines into the hip muscles. Each batch of vaccines will be injected with 5 piglets, each with 2ml, and observe the clinical manifestations and measure the body temperature.

[0079] 2) The prepared vaccines were injected into healthy sows on the 70th day of gestation through separate injections into the hip muscles. Each batch of vaccines were injected with 5, 2ml each, and the clinical manifestations and farrowing conditions were observed.

[0080] 2. Test results

[0081] 3.2.1 The safety of the subunit vaccine of the present invention for vaccination of weaned piglets

[0082] After the prepared vaccine was inoculated with 35-day-old healthy we...

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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. technical background [0002] Streptococcus suis (S. suis) is the main pathogen causing swine streptococcus. According to the capsular polysaccharide (CPS) on the bacterial surface, S. suis can be divided into 33 serotypes, namely type 1 / 2, Type 1~31 and 33. Among them, Streptococcus suis 2 (SS2) is the serotype with the strongest virulence, widespread prevalence and the highest frequency of clinical isolation. SS2 is an important zoonotic pathogen, which can cause porcine meningitis, endocarditis, sepsis, arthritis, serositis, pneumonia, etc. It often causes the sudden death of weaned piglets or fattening pigs; humans are infected with this bacteria, It can cause meningitis, permanent deafness, sepsis, endocarditis, and even death. In 1991, it was first reported that a suspected case of swine t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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