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Coiled-coil domain containing protein 8 (CCDC8) having inhibition on HIV-1 and application thereof

A coiled-coil structure, HIV-1 technology, applied in the field of biomedicine, can solve the side effects of HIV, can not completely cure AIDS, drug resistance and other problems

Active Publication Date: 2015-11-11
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to solve the toxic and side effects, drug resistance, and compliance problems caused by the existing drugs for treating AIDS, and the problem that AIDS cannot be completely cured, and to provide a protein coiled-coil structure encoded by human cells that strongly inhibits HIV-1 Protein 8 and its application

Method used

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  • Coiled-coil domain containing protein 8 (CCDC8) having inhibition on HIV-1 and application thereof
  • Coiled-coil domain containing protein 8 (CCDC8) having inhibition on HIV-1 and application thereof
  • Coiled-coil domain containing protein 8 (CCDC8) having inhibition on HIV-1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1, HIV-1 resistance experiment of CCDC8

[0025] First, the plasmid pTT5-SH5-CCDC8 expressing CCDC8 was constructed. The vector pTT5-SH5 was constructed by Yves Durocher, Biotechnology Research Institute, Montreal, Canada and the vector sequence was provided. Using the method of reverse transcription polymerase chain reaction (RT-PCR), the primers are TT-CCDC8-BamHI and TT-CCDC8-EcoRI (as shown in Table 1) to amplify CCDC8, and the amplified product is treated with restriction endonuclease After digestion with BamHI and EcoRI, clone into pTT5-SH5 vector. The CCDC8 sequence was determined and confirmed by Invitrogen (SEQIDNo.1), and the protein sequence was translated according to the triple codon rule (SEQIDNo2).

[0026] Then, HEK293T cells (CRL-11268, ATCC) were cultured in DMEM medium supplemented with 10% serum and penicillin in a 37°C incubator containing 5% carbon dioxide. After recovery, the cells are cultured for two weeks and can be used after being ...

Embodiment 2

[0034] Example 2, CCDC8 can cause polyubiquitination and degradation of HIV-1 Gag

[0035] CCDC8 interacts with Gag, OBSL1 and Cul7. Checking the literature shows that CCDC8 is related to 3M syndrome, a genetic disease. 3M syndrome is an autosomal recessive genetic disease, mainly characterized by short stature but normal intelligence. Using exome sequencing, 70% of patients had mutations in the E3 ligase Cul7, 25% had mutations in OBSL1, and 5% had mutations in CCDC8. 3M syndrome and in vitro co-immunoprecipitation experiments linked CCDC8, OBSL1 and Cul7 together, suggesting that they are in a cellular pathway. We also found that OBSL1 or Cul7 alone had no anti-HIV-1 effect ( Image 6 A). Our co-immunoprecipitation experiments also showed that HIV-1Gag interacted with CCDC8 ( Image 6 B), Cul7 also interacts with CCDC8 and OBSL1 ( Image 6 C). The specific method is as follows. After 48 hours of transfection, the cells were lysed with the following buffer solution (20...

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Abstract

The invention relates to new coiled-coil domain containing protein 8 (CCDC8) having inhibition on HIV-1 and an application thereof. The CCDC8, which is generated by human cells, in the invention has a strong anti-HIV-1 effect. By means of combination of the CCDC8 with matrix protein region of HIV-1Gag, assembly of the Gag protein on cytomembrane is inhibited. The CCDC8, OBSL1, and E3 ubiquitin ligase Cul7 cause internalization, multi-ubiquitination and degradation of the HIV-1Gag. According to the invention, on the basis of the CCDC8, a new plasmid is established and can be used for express new proteins, namely, the protein composed of 274 amino acids at the N-terminal of the CCDC8, thereby degrading HIV-1.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a novel protein capable of inhibiting HIV-1. Specifically, the anti-HIV-1 effect can be achieved by inducing polyubiquitination and degradation of HIV-1 Gag. Background technique [0002] AIDS (AcquiredImmuneDeficiencySyndrome, AIDS) caused by Human Immunodeficiency Virus (Human ImmunodeficiencyVirus, HIV-1) or AIDS virus has been prevalent in the world for more than 30 years. AIDS is a severe infectious disease. The virus directly attacks the immune system of the human body, resulting in defective immune function of the human body, prone to opportunistic infections and tumors, and most of them die. According to the report of the World Health Organization (WHO), by the end of 2013, there were about 35 million HIV-1 living infected people in the world, and about 30 million people had died, totaling about 65 million people. In 2013, about 2.1 million people were newly infected with HIV-1, ...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/47A61K38/17A61P31/18
Inventor 魏民邵一鸣劳闰斯·克雷曼
Owner NANKAI UNIV
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