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Construction and application of SHIV (simian/human immunodeficiency virus) infectious clone

An infectious clone, SHIV-KB9 technology, applied in the construction and application of SHIV infectious clones, can solve the problem of not being able to simulate the dynamic changes of host lymphocytes well

Inactive Publication Date: 2015-03-11
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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Problems solved by technology

After infection of host cells by these SHIV viruses, CD4+ lymphocytes will continue to decline, showing more characteristics of late HIV-1 infection, which cannot well simulate the symptoms caused by natural HIV-1 infection. Dynamics of host lymphocyte depletion

Method used

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  • Construction and application of SHIV (simian/human immunodeficiency virus) infectious clone
  • Construction and application of SHIV (simian/human immunodeficiency virus) infectious clone
  • Construction and application of SHIV (simian/human immunodeficiency virus) infectious clone

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Embodiment Construction

[0078] The construction and application of the SHIV infectious clone provided by the present invention, first clone the full-length genome of the highly pathogenic and virulent strain SHIV-KB9, which is widely used in the world, into a low copy number On the vector pBR322, a highly stable plasmid SHIV-KB9 / pBR322 was obtained. Then, using SHIV-KB9 / pBR322 as the genome backbone, the env gene of CRF08_BC, the main epidemic strain of HIV-1 in China, was used to replace its corresponding gene fragment, and an infectious clone SHIV-KBQJ-12 of the chimeric CRF08_BC env gene was obtained. The newly substituted env gene is derived from HIV-1CRF08_BC infectious clone QJ001, including the entire intracellular region, transmembrane region and part of the extracellular region of gp120 and gp41, and the position of the substituted env gene corresponds to the standard reference The nucleotide position of strain HXB2 is 6103-8249; the invention is easy to operate, has a high success rate and ...

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Abstract

The invention relates to construction and application of SHIV (simian / human immunodeficiency virus) infectious clone. Firstly, the overall length genome of a virulent strain SHIV-KB9 embedded with a B subtype env gene is cloned to a low copy number carrier pBR322, the obtained plasmid SHIV-KB9 / pBR322 is taken as a genome skeleton, the env gene of Chinese main prevalent strain CRF08_BC of HIV-1 is used for substituting for corresponding gene segments so as to obtain the infectious clone SHIV-KBQJ-12, the replacing env gene is from the infectious clone strain QJ001 of HIV-1CRF08_BC and comprises the whole intracellular region, a transmembrane region and part of an extracellular region of gp120 and gp41, and the position of the replacing env gene, corresponding to the position of the nucleotide of a standard reference strain HXB2, is 6103-8249. The construction is simple and convenient to operate, has high success rate, can obtain the infectious clone in a short time, and can be used for establishing an SHIV / Chinese rhesus infection model.

Description

technical field [0001] The invention relates to the technical field of gene recombination, in particular to a novel chimeric monkey / human immunodeficiency virus ( A method for constructing an infectious clone of Chimeric simian / human immunodeficiency virus (SHIV), and also involves using the SHIV infectious clone to establish a SHIV / rhesus infection model. [0002] The present invention is based on the research results of the molecular clone SHIV-KB9 of the highly pathogenic and virulent strain SHIV-89.6P widely used in the world and the infectious molecular clone QJ001 of HIV-1CRF08_BC constructed in our laboratory. Among them, the 3'-SHIV-KB9 and 5'-SHIV-KB9 plasmids provided by the HIV / AIDS Reagent Program can amplify the full-length genome of SHIV-KB9; the infectious molecular clone QJ001 is preserved in Escherichia coli STBL3, which The bacillus was named STBL3 / QJ001 (Escherichia coli STBL3 / QJ001), and it was preserved by the China Center for Type Culture Collection (Add...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/63A01K67/027C12R1/93
Inventor 杨荣阁黄黎草川茂孙宾莲
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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