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Application of protein Rh054_01780 to rickettsia heilongjiangensis-resistant immune protection

A rickettsia and protein technology, applied in the direction of resistance to vector-borne diseases, antibacterial drugs, bacterial antigen components, etc., can solve the problems of high cost, complicated process, and high protection requirements

Inactive Publication Date: 2012-10-10
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

[0004] The usual method of preparing spotted fever vaccine is to use inactivated bacteria or chemically extract subunit vaccines, and these methods require a large number of chicken embryos or cells to culture Rickettsia, but the protection requirements for extracting and purifying Rickettsia are high, the process is complicated, and the cost is high. Expensive, so it is difficult to prepare and promote the use of large quantities

Method used

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  • Application of protein Rh054_01780 to rickettsia heilongjiangensis-resistant immune protection
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  • Application of protein Rh054_01780 to rickettsia heilongjiangensis-resistant immune protection

Examples

Experimental program
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Effect test

Embodiment 1

[0037] Embodiment 1, preparation of recombinant plasmid and protein

[0038] 1. Construction of recombinant plasmids

[0039] 1. Construction of recombinant plasmid A

[0040] (1) Genomic DNA of Rickettsia heilongjiang was extracted.

[0041] (2) Using the genomic DNA extracted in step (1) as a template, perform PCR amplification with a primer pair composed of F1 and R1 to obtain a PCR amplification product.

[0042] F1: 5'-GT GGATCC GGAGAAAGCGAAACTA-3';

[0043] R1:5'-AT CTCGAG CTTGGATGCAGCAGTT-3'.

[0044] The target sequence of F1 and R1 is the double-stranded DNA molecule shown in sequence 3 (coding the protein Rh054_01780 shown in sequence 1 in the sequence listing).

[0045] PCR amplification conditions: 94°C pre-denaturation for 5 minutes; 35 cycles of 95°C for 30 sec, 55°C for 30 sec, and 72°C for 1.5 min; 72°C for 7 min.

[0046] (3) The PCR amplified product of step (2) was double-digested with restriction endonucleases BamHI and XhoI, and the digested produ...

Embodiment 2

[0073] Example 2, Protective Evaluation of Antigenic Protein Rh054_01780

[0074] Twenty male C3H / HeN mice aged 3-4 weeks were randomly divided into four groups, with 5 mice in each group, and were treated in parallel as follows (the following immunization doses refer to the total volume of immunization solution and Freund's adjuvant) :

[0075] The first group (Rh054_01780 group): Take the protein A solution prepared in Example 1, adjust the protein concentration to 0.6mg / ml with the elution buffer, and use it as the solution for immunization; on the 0th day, mix the solution for immunization with Freund's complete Mix equal volumes of immunization solution and Freund's incomplete adjuvant in equal volumes and prepare an emulsion by ultrasonic for the first immunization (immunization dose: 100 μl / monkey). Emulsion, for the second immunization (immunization dose: 100 μl / monkey); on the 42nd day, mix the immunization solution with Freund’s incomplete adjuvant in equal volumes ...

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Abstract

The invention discloses application of protein Rh054_01780 to rickettsia heilongjiangensis-resistant immune protection, and provides application of the protein Rh054_01780 to preparation of a vaccine for far eastern spotted fever, a vaccine for rickettsia heilongjiangensis, a protective antigen for far eastern spotted fever, or a protective antigen for rickettsia heilongjiangensis. The protein Rh054_01780 is (a) or (b), wherein (a) is a protein which has an amino acid sequence shown as a sequence 1 in a sequence table; and (b) is a protein which is formed by substituting and / or deleting and / or adding one or more amino acid residues in the amino acid sequence shown as the sequence 1 in the sequence table, has the same function and is derived from the sequence 1. Compared with a mycoproteinvaccine, a vaccine prepared from the protein Rh054_01780 has the advantages of simple and convenient preparation method, low cost, high yield, safety and the like; and the protein Rh054_01780 has significant value in the aspect of epidemic prevention and treatment of far eastern spotted fever.

Description

technical field [0001] The invention relates to the application of protein Rh054_01780 in the immune protection against Rickettsia heilongjiang. Background technique [0002] Rickettsial disease is a zoonotic natural foci disease that seriously threatens human health. It brought serious disasters to humans during the two world wars. Spotted fever is a type of rickettsial disease, caused by the spotted fever group Rickettsia, and is an infectious disease transmitted by ticks. [0003] Rickettsia heilongjiangensis is a new Rickettsia strain isolated from forest leather ticks in Suifenhe area, Heilongjiang, my country in 1983. It is an obligate intracellular parasitic Gram-negative small bacillus. The spotted fever disease caused by this bacterium was officially named Far Eastern Spotted Fever by WHO. Far Eastern spotted fever is a zoonotic disease that is mainly distributed in Northeast China, Siberia in Russia and Japan, and has also been found in Guangdong in recent years....

Claims

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Application Information

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IPC IPC(8): A61K39/02A61P31/04
CPCY02A50/30
Inventor 齐永温博海王锡乐熊小路段长松李佳明焦俊龚文平贾引军
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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