Single-circulation-copy AIDS virus-like particle and preparation method and application thereof

An HIV, single-cycle technology, applied in the direction of biochemical equipment and methods, viruses, antiviral agents, etc., to achieve the effect of easy observation and tracking, high sensitivity and high safety

Inactive Publication Date: 2016-07-13
CENT SOUTH UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the high pathogenicity of the HIV-1 virus, the precautionary facilities required to study various anti-HIV strategies must reach Biosafety Level 3 (BSL-3), and the above-mentioned experiments on HIV infection on humanized mice are also the same. It needs to be carried out in a three-level biosafety laboratory. Many scientific research teams have made great contributions to the fight against HIV, but because of the need for high-level security facilities, they are restricted from further research

Method used

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  • Single-circulation-copy AIDS virus-like particle and preparation method and application thereof
  • Single-circulation-copy AIDS virus-like particle and preparation method and application thereof
  • Single-circulation-copy AIDS virus-like particle and preparation method and application thereof

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Embodiment

[0034] Experimental group: the production of a single-cycle replication HIV-like particle (C-ΔRI / ΔE / Glu), firstly delete the reverse transcriptase, integrase and part of the membrane glycoprotein gene sequence of the HIV proviral plasmid, and at the same time use the Gaussialuciferase gene Substituting the viral nef gene sequence, the proviral structure produced in this way is named as ΔRI / ΔE / Gluc plasmid, as the base sequence described in SEQ ID NO.1 in the sequence listing. Then the ΔRI / ΔE / Gluc plasmid, CMV-Gag / Pol expression plasmid, and viral envelope glycoprotein Env expression plasmid were co-transfected into isolated human 293T cells to complement the reverse transcriptase, integrase and Envelope defect, thus screened and isolated single-cycle replicating HIV-like particles, namely C-ΔRI / ΔE / Gluc. The constructs of the ΔRI / ΔE / Gluc plasmid, the CMV-Gag / Pol expression plasmid, and the viral envelope glycoprotein Env expression plasmid are as follows: figure 1 As shown in ...

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Abstract

The invention discloses single-circulation-copy AIDS virus-like particle and a preparation method and application thereof.The virus-like particle is obtained through separation and screening after an HIV provirus plasmid with multiple genes being deleted, a CMV-Gag / Pol expression plasmid and an Env expression plasmid co-transfect cells, and the HIV provirus plasmid has the base sequence as shown in SEQ ID No.1 after the HIV provirus plasmid replaces the virus nef gene sequence for marking through a Gaussia luciferase gene sequence.The virus-like particle is used for preparing a monoclonal antibody using immune animals for diagnosing and treating, a targeting vaccine serving as a dendritic cell, and the infection source for CD4+T cell system in-vitro infection and animal AIDS virus infection.The single-circulation-copy AIDS virus-like particle is high in sensitivity, high in safety, and applicable to different labs without three-level lab closed environments.

Description

technical field [0001] The invention belongs to the technical field of virus-like particles, and in particular relates to a single-circulation replication HIV-like particle and its preparation method and application. Background technique [0002] Acquired Immunodeficiency Syndrome (AIDS) is a degenerative disease of the immune and nervous systems caused by HIV-1 infection. This infectious disease has threatened the survival, welfare and social stability of the world. Global estimates of HIV epidemiology indicate that in 2012, there were already 35.3 million HIV-1 carriers worldwide, with 2.3 million new cases that year alone. Even with the remarkable progress in the field of anti-HIV-1 chemotherapy over the past few decades, effective prevention of HIV-1 infection remains one of the most important challenges due to the rapid emergence of viral escape strains and drug resistance. Therefore, developing novel anti-HIV methods and better understanding the mechanism of latent HI...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/04C07K16/10A61K39/21A61K49/00A61P31/18
CPCC12N7/00A61K39/00A61K49/0004C07K16/1045C12N2740/16023
Inventor 姚小剑姚小刚敖竹君欧阳清检
Owner CENT SOUTH UNIV
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