Recombinant tubercle bacillus antigen poly epitope peptide and its constructing method and use

A Mycobacterium tuberculosis, multi-epitope technology, applied in the field of molecular immunology, can solve problems such as death, and achieve the effect of promoting the solution and promotion of multi-drug resistance problems

Inactive Publication Date: 2007-01-24
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Claims
  • Application Information

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Problems solved by technology

Intravenous injection of Mycobacterium tuberculosis into β2m-deficient mice can lead to rap...

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  • Recombinant tubercle bacillus antigen poly epitope peptide and its constructing method and use
  • Recombinant tubercle bacillus antigen poly epitope peptide and its constructing method and use
  • Recombinant tubercle bacillus antigen poly epitope peptide and its constructing method and use

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Embodiment 1

[0037] 1 Materials and methods

[0038] 1.1 Research object

[0039] 10 HLA-A*0201-positive healthy volunteers and 6 HLA-A*0201-negative healthy volunteers were all PPD+. 20ml of peripheral blood was collected and anticoagulated with heparin.

[0040] 1.2 Plasmids, strains and reagents

[0041] Plasmids pET22b and pGEX-4T-1, Escherichia coli DH5α and BL21 strains were purchased from Novagen

[0042] PCR kit: Promega

[0043] Plasmid mini-extraction kit, mini-gel recovery kit: Youjing Bioengineering Co., Ltd.

[0044] Restriction enzymes (BamHI, NdeI, XhoI): Takara

[0045] T 4 DNA ligase: Roche

[0046] Agarose: Gibco

[0047] Tryptone, yeast extract: Oxoid

[0048] Isopropylthio-β-D-galactoside (IPTG): Shanghai Boguang Biotechnology Co., Ltd.

[0049] Nitrocellulose membrane: Amersham

[0050] Alkaline phosphatase (ACP)-labeled goat anti-human IgA: KPL company

[0051] Pre-stained protein marker: bio-lab company

[0052] Lymphoprep TM Purchased from Norway AXIS-...

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Abstract

The present invention belongs to the field of molecular immunology, discloses six antigen epitope molecule simulating peptides capable of exciting human body's tubercle bacillus antagonizing protective immunoreaction, and provides recombinant tubercle bacillus antigen polyepitope peptides, which are fusion peptides with the amino acid sequences as shown in SEQ ID No.1, SEQ ID No. 2, SEQ ID No. 3, SEQ ID No. 4, SEQ ID No. 5 and SEQ ID No. 6 separately. The present invention also provides their construction process and use. These polyepitope peptides, cell and animal experiments show, can excite efficient specific tubercle bacillus antagonizing protective immune response. The present invention may be used in preventing and treating tuberculosis.

Description

technical field [0001] The invention relates to the field of molecular immunology, in particular to a recombinant Mycobacterium tuberculosis antigen multi-epitope peptide capable of stimulating human body's protective immune response against Mycobacterium tuberculosis and its construction method and application. Background technique: [0002] Tuberculosis (TB) is one of the main killers threatening human health around the world. About 2 billion of the world's population has been infected with Mycobacterium tuberculosis (Mtb), and the vast majority of infected people have no symptoms and become latent infection (latentMtb.infection, LTBI), of which about 10% of latent infections can evolve into Tuberculosis, of which 16 million are active. Worldwide, about 8 million people are newly infected each year, and about 2 million people die from tuberculosis. In China, in the national infectious disease epidemic report released by the Ministry of Health in November 2005, the incide...

Claims

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Application Information

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IPC IPC(8): C07K14/195C12N15/62C12N15/31A61K39/04A61P31/06
Inventor 邓安梅仲人前娄加陶吴传勇陈孙孝周晔陈燕陈波蒋天舒谷明莉
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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