Anti-HIV aedes albopictus protein with platelet aggregation inhibiting function and preparation method of anti-HIV aedes albopictus protein

A technology of platelet aggregation and Aedes albopictus, applied in the field of protein obtained from animal tissues, can solve the problem of insufficient research on pharmacologically active components

Inactive Publication Date: 2015-02-18
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] my country has a long history of medicinal research on animal toxins, but there are few

Method used

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  • Anti-HIV aedes albopictus protein with platelet aggregation inhibiting function and preparation method of anti-HIV aedes albopictus protein
  • Anti-HIV aedes albopictus protein with platelet aggregation inhibiting function and preparation method of anti-HIV aedes albopictus protein
  • Anti-HIV aedes albopictus protein with platelet aggregation inhibiting function and preparation method of anti-HIV aedes albopictus protein

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Embodiment Construction

[0018] Preparation and identification of anti-HIV Aedes albopictus protein with platelet aggregation inhibitory function:

[0019] 1) Cloning of anti-HIV Aedes albopictus protein gene with platelet aggregation inhibiting function:

[0020] I. Extraction of total RNA from Aedes albopictus salivary gland:

[0021] A. live Aedes albopictus is placed on ice for 30-60 minutes and frozen for anesthesia, dissects 50 Aedes albopictus under the microscope to obtain salivary glands, adds 1m1 total RNA extraction buffer (Trizol solution, the product of U.S. GIBCOBRL company), in 5m1 Homogenize for 1 min in a glass homogenizer.

[0022] B. Add an equal volume of phenol / chloroform solution, mix vigorously, leave at room temperature for 1 minute, centrifuge at 12,000 rpm for 10 minutes at 4°C, and discard the precipitate.

[0023] C. Add an equal volume of isopropanol to the supernatant, let it stand at room temperature for 4 minutes, centrifuge at 12000 rpm for 10 minutes at 4°C, wash th...

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Abstract

The invention relates to an anti-HIV aedes albopictus protein with a platelet aggregation inhibiting function. An amino acid sequence of the anti-HIV aedes albopictus protein is shown as SEQ ID NO:1. A preparation method of the anti-HIV aedes albopictus protein comprises the following steps: purifying total RNA of aedes albopictus salivary glands and amplifying so as to obtain total cDNA, then amplifying genes of the total cDNA taken as a template and cloning the genes into a prokaryotic expression vector, carrying out inducible expression via IPTG, carrying out denaturation and renaturation folding on an expressed inclusion body, then filtering the inclusion body via a gel, and carrying out separation and purification on the inclusion body by ion exchange chromatography so as to obtain the anti-HIV aedes albopictus protein. The anti-HIV aedes albopictus protein has the activities of inhibiting the proliferation of the HIV in cells and the platelet aggregation induced by multiple agonists, and has the advantages of no cytotoxicity and strong anti-HIV specificity. Thus, the anti-HIV aedes albopictus protein can be used for preparing medicines for resisting AIDS (Acquired Immune Deficiency Syndrome) and related diseases.

Description

Technical field: [0001] The present invention relates to peptides with more than 20 amino acids, in particular to a protein obtained from animal tissue, which has biological activity against HIV. Background technique: [0002] AIDS (AIDS) is an infectious disease characterized by severe damage to the systemic immune system. Since its discovery, it has seriously threatened the survival of human beings and has had a huge impact on the health of the global population and the development of social economy. With years of research, there are many anti-AIDS drugs. According to the different targets of drugs acting on the virus, the anti-AIDS drugs that have been marketed can be roughly divided into glycoside reverse transcriptase inhibitors, non-nucleoside reverse transcriptase inhibitors, protease inhibitors, entry inhibitors, integrase inhibitors, etc. . These drugs respectively inhibit the processes of entry, fusion, reverse transcription, integration, transcription, translati...

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Application Information

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IPC IPC(8): C07K14/435C12N15/70C07K1/18C07K1/16A61P31/18
CPCC07K14/43563
Inventor 徐学清李琳刘叔文
Owner SOUTHERN MEDICAL UNIVERSITY
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