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HIV-1 multi-epitope recombinant protein, and encoding gene and application thereof

A HIV-1 coding technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problem of no vaccine application, etc., and achieve good safety, strong immunogenicity, and good immunogenicity

Active Publication Date: 2014-11-26
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

Vaccine is an effective means to control HIV infection. Although important progress has been made in the research and development of HIV vaccine, a phase III clinical trial of HIV vaccine RV144, which can induce humoral and cellular immunity at the same time, shows that the vaccine can reduce HIV infection to a certain extent. infection to reduce the incidence of AIDS, but there is still no effective vaccine for the population

Method used

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  • HIV-1 multi-epitope recombinant protein, and encoding gene and application thereof
  • HIV-1 multi-epitope recombinant protein, and encoding gene and application thereof
  • HIV-1 multi-epitope recombinant protein, and encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1, Expression and purification of HIV-1 multi-epitope recombinant protein HIV-MEP1

[0054] 1. Conserved epitope screening and DNA sequence synthesis in line with the restriction of dominant MHC in the Chinese population

[0055] Obtain all reported epitopes on the env, gag and pol genes of the main HIV-1 epidemic strains in China (CRF01_AE, CRF07_BC, CRF08_BC, B) from HIV Databases, and screen out all epitopes that are conserved in sequence and conform to the MHC-restricted dominant Chinese population , the epitopes screened were further connected in series with the linker sequence GGGS and codon-optimized in Escherichia coli, and the corresponding double-stranded DNA was synthesized in Nanjing GenScript Company (and BamH I and Hind III recognition sequences were added at both ends), as shown in No. 31-499 of Sequence 2 in the Sequence Listing (No. 31-36 is the BamH I recognition sequence, No. 494-499 is the Hind III recognition sequence, and No. 493-495 is the...

Embodiment 2

[0067]Example 2, Immunological evaluation of HIV-1 multi-epitope recombinant protein HIV-MEP1 in BALB / c mice

[0068] 1. Animal immunization and specimen collection:

[0069] Female BALB / c mice (6-8 weeks old) were randomly divided into three groups, i.e. recombinant protein aluminum adjuvant group (group 1), recombinant protein alone immunization group (group 2) and PBS control group (group 3), A total of three immunizations were performed, with the time interval of each immunization being 4 weeks, and the dose of each immunization was the same. The grouping situation and immunization strategy are shown in Table 1.

[0070] Table 1 Animal immunization experiment grouping and immunization strategy

[0071] group

quantity

Immunoreagent

immune dose

way of immunization

1

6

Aluminum Adjuvant+HIV-MEP1 / PBS

10μg / 200μl

intramuscular injection

2

6

HIV-MEP1 / PBS

10μg / 200μl

intramuscular injection

3

5

...

Embodiment 3

[0101] Example 3. Immunological evaluation of HIV-1 multi-epitope recombinant protein HIV-MEP1 in human HLA A2.1 / DR1 transgenic mice

[0102] The experimental scheme of this embodiment is the same as that of Example 2, except that human HLA A2.1 / DR1 transgenic mice are used to evaluate cellular immunity (IFNγ), since human HLA A2.1 / DR1 transgenic mice are C57 / B6 Background transgenic mice, in order to evaluate whether the cellular immunity induced by HIV-1 multi-epitope recombinant protein HIV-MEP1 is restricted by human MHC, at the same time, C57 / B6 mice were used as control for comparative study. The inoculation route, inoculation dose, and evaluation method are the same as in Example 2.

[0103] The results show:

[0104] (1) Stimulation results of HIV-1 multi-epitope recombinant protein HIV-MEP1 and 12 mixed polypeptides

[0105] Spleen cells of C57 / B6 mice were immunized with HIV-1 multi-epitope recombinant protein HIV-MEP1 and mixed polypeptides to stimulate aluminum a...

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Abstract

The invention discloses an HIV-1 multi-epitope recombinant protein, and an encoding gene and an application thereof. The protein provided by the invention is as follows: a) protein formed by amino acid sequences shown in the 13th to 164th bits of a sequence 1; b) protein formed by amino acid sequences shown in the sequence 1; or c) protein, which is obtained from the protein defined in a) or b) in a manner of replacing and / or missing and / or adding of one or more amino acid residues, and is related to anti-HIV-1. An experiment proves that the HIV-1 multi-epitope recombinant protein disclosed by the invention has good immunogenicity, and is an immunoreactive protein which has an application prospect and can be applied to prevention and treatment of HIV-1.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to an HIV-1 multi-epitope recombinant protein and its encoding gene and application. Background technique [0002] AIDS is acquired immunodeficiency syndrome (acquired immunodeficiency syndrome, AIDS) caused by human immunodeficiency virus (human immunodeficiency virus, HIV) infection. Since the first AIDS case was reported in the world in 1981, AIDS has spread rapidly around the world in just over 30 years, becoming the biggest health crisis in the world today and an important public health issue affecting public health in my country. In recent years, there has been a new epidemic trend of AIDS infection in my country, that is, the incidence rate is increasing year by year, the infected objects cover different groups of people, the proportion of sexual transmission is increasing, especially the increase in male-gay male patients, and the proportion of mutated recombinant viruses in the e...

Claims

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Application Information

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IPC IPC(8): C07K14/16A61K39/21A61P31/18C12N15/49C12N15/70
CPCA61K39/00C07K14/005C12N2740/16022C12N2740/16034
Inventor 周育森寇志华杨裔郭彦于虹孙世惠赵光宇李军锋
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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