Staphylococcus aureus fusion protein of fibronectin and bindin, its preparation method and uses

A Staphylococcus and fusion protein technology, applied in the field of fusion proteins, can solve the problems that have not yet been reported on the fusion protein of fibronectin binding protein FnBP and Trx, and achieve the effect of improving the protective effect and enhancing the immunogenicity

Inactive Publication Date: 2006-12-27
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] There is no report on the fusion protein of fibronectin-binding protein FnBP and Trx

Method used

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  • Staphylococcus aureus fusion protein of fibronectin and bindin, its preparation method and uses
  • Staphylococcus aureus fusion protein of fibronectin and bindin, its preparation method and uses
  • Staphylococcus aureus fusion protein of fibronectin and bindin, its preparation method and uses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment one Staphylococcus aureus surface protein fnb gene is in

[0037] Cloning, expression and purification in Escherichia coli

[0038] 1. Materials

[0039] 1. Bacterial strain: Staphylococcus aureus (S.aureus) FRI 1169 is an international standard strain and is commercially available.

[0040] 2. Vector and host bacteria: The cloning vector PMD 18-T Vector is the product of Takara, the expression vector pET-32a(+)Vecter is the product of Novagen, Escherichia coli DH5α, and Escherichia coli BL21(DE3) are the products of Stratagene.

[0041] 3. Culture medium: LB medium, prepared according to conventional methods.

[0042] 4. Tool enzymes: endonuclease BamH I, Nde I, Hind III, Xhol I; ligase Ligation Solution I are all products of Takara Company.

[0043] 5. Main reagents and kits: PCR reagents: purchased from Shanghai Sangon, X-gal, IPTG, DNAMarker DL2000 is a product of Takara Company, imidazole is a product of QIAGEN, sodium dod...

Embodiment 2

[0100] Example 2 Study on the immune effect of Trx-FnBP protein

[0101] 1. Materials

[0102] 1. Recombinant protein: Trx-FnBP protein is a low-temperature freeze-dried product after dialysis of the purified protein.

[0103] 2. Immune adjuvants: FREUND’S ADJUVANT Complete and FREUND’SADJUVANT Incomplete are products of SIGMA.

[0104] 3. Other reagents: MUCIN (Type II: Crude; From Porcine Stomach) is a product of SIGMA; Ficoll-paque is a product of Pharmacia; CellTiter 96 R Aqueous Non-RadioactiveCell Proliferation Assay is a kit from Promega; Thioglycolate Broth with Resazurine is a product from Fluka; goat anti-mouse IgG-HRP is a product from Beijing Zhongshan Biotechnology Company; ELISA reagents are all analytically pure.

[0105] 4. Experimental animals: Kunming secondary female mice and Balb / c female mice at 5-6 weeks were purchased from the Experimental Animal Center of the Academy of Military Medical Sciences.

[0106] 2. Experimental method

[0107]...

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Abstract

The invention discloses a fusion protein for Micrococcus pyogenes surface protein, and also discloses the method for preparing the same as well as its usage. The fusion protein is fused by Micrococcus pyogenes fnb and thioredoxin Trx on expression carrier, and the process comprises preparation of Micrococcus pyogenes surface protein fnb coding gene, colony of fnb coding gene into expression carrier and protein fusion. The fusion protein is soluble protein and retains immunity of natural protein. It is detected that said fusion protein is characterized by good response for humoral immunity and cell immunity, enlargement of protection range of protein vaccine and prevention of infection caused by most of Micrococcus pyogenes.

Description

technical field [0001] The invention relates to a fusion protein, in particular to a fusion protein of Staphylococcus aureus fibronectin binding protein, and also relates to a preparation method and application of the fusion protein. Background technique [0002] Staphylococcus aureus is the most common pathogenic bacteria in nosocomial infections, and it is also the main pathogenic bacteria of large-scale infections in the society. Staphylococcus aureus infection can cause diseases such as endocarditis, osteomyelitis, septic arthritis, pneumonia and abscess. Staphylococcus aureus is also the main pathogen of important economic animals. The main problem caused by S. aureus resistance to first-line drugs such as penicillin is that methicillin-resistant S. aureus strains are resistant to most clinically used antibiotics, so about 10-60% of S. aureus were isolated from hospitals; of particular concern are the increasing susceptibility of MRSA strains to vancomycin, the antibi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/63A61K39/085A61P31/04A61P11/00A61P19/02A61P19/08A61P9/00
Inventor 姜永强郑玉玲周宏宁保安马茹李韩平
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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