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Borrelia garinii OspA protein C-terminal peptide fragment and application thereof

A technology of Borrelia garzii and its C-terminus, which is applied in the field of immunology, can solve problems such as unseen related researches, and achieve good immune protection effect

Active Publication Date: 2019-11-22
ICDC CHINA CDC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, studies have proved that the OspA full-length protein of the main pathogenic genotypes of B. garinii and B. afzelii in China has good immune protection for animals, but removal may cause side effects. Whether the OspA peptide after the reaction fragment can produce the same immune protection effect on the body as the full-length protein has not yet been researched

Method used

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  • Borrelia garinii OspA protein C-terminal peptide fragment and application thereof
  • Borrelia garinii OspA protein C-terminal peptide fragment and application thereof
  • Borrelia garinii OspA protein C-terminal peptide fragment and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Example 1 Expression and identification of the C-terminal peptide of Borrelia garzii OspA protein

[0039] 1. Cloning and expression of OspA gene fragment

[0040] The gene fragment of the outer membrane protein OspA 126-274aa of Chinese Lyme spirochete B.garinii genotype PD91 (that is, the cross-reactive sequence has been removed from the full-length sequence of the OspA protein), with EcoRI and HindIII restriction sites added at both ends, and Cloned and expressed after being fused with the His tag, the nucleic acid sequence of the OspA gene fragment is shown in SEQ ID NO:2.

[0041] The primers used are as follows:

[0042] Upstream primer: 5'- GAATTC TTTAATGCAAAAGGTGA-3' (underlined as EcoRI restriction site);

[0043] Downstream primer: 5'- AAGCTT TTATTTTAAAGCTGTTTTAAG-3' (underlined HindIII restriction site).

[0044]The OspA gene fragment and the plasmid PET-30a were subjected to a double digestion reaction, and the two double digestion products were ligat...

Embodiment 2

[0049] Example 2 Immunoprotective study of rOspA-pep protein

[0050] 1. Screening of optimal immune dose of rOspA-pep protein

[0051] Set up 7 dose groups (20, 30, 40, 50, 60, 80, 100 μg) and 1 blank control group. Dosage groups were immunized with New Zealand rabbits (5 rabbits in each group) with different doses of rOspA-pep protein, and the control group was injected with the same volume of PBS to screen the optimal immune dose of rOspA-pep. The IgG antibody titer of rabbit serum was detected before immunization and on the 15th, 30th, 45th and 60th day after immunization by indirect immunofluorescence (IFA) method. The rabbit serum of the dose group with higher titer was selected for further neutralization test. Select 10 New Zealand rabbits (body weight 2.5kg, half male and half male) to be immunized with rOspA-pep protein at the optimal dose, and the other 5 rabbits were used as blank control group (the control group was injected with PBS solution). The first immuniz...

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Abstract

The invention provides a Borrelia garinii OspA protein C-terminal peptide fragment and an application thereof. The OspA peptide fragment (126-274aa), containing the His label, of the Borrelia gariniiprovided by the invention has good immune protection on rabbits, and does not generate adverse side effects on organisms. The invention provides an effective means for treating or preventing Borreliaburgdorferi infection.

Description

technical field [0001] The invention relates to the field of immunology, in particular to the C-terminal peptide of Borrelia garzii OspA protein and its application. Background technique [0002] Lyme disease is a natural foci disease. In nature, Borrelia burgdorferi infects ticks and host small mammals. When infected ticks bite humans, humans can develop Lyme disease. Mainly cause flu-like symptoms, which may or may not be accompanied by chronic migratory erythema, arthritis, neurological disease, myocarditis and other symptoms: chronic atrophic extremity dermatitis, conjunctivitis, optic neuritis, etc. Because the clinical manifestations of patients with Lyme disease are complex and diverse, and there is currently no unified diagnostic standard for Lyme disease in China, various factors lead to misdiagnosis of patients and delayed treatment. [0003] Vaccine development is an effective way to control the spread of Lyme disease in the population, but no effective Lyme dise...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/20C12N15/31A61K39/02A61P31/04
CPCA61K39/0225A61P31/04C07K14/20Y02A50/30
Inventor 郝琴苗广青张琳侯学霞
Owner ICDC CHINA CDC
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