HIV medicament screening cell model and special pseudotype lentivirus therefor

A cell model, lentivirus technology, applied to cells modified by introducing foreign genetic material, viruses/phages, applications, etc., can solve the problems of low sensitivity, complicated result detection procedures, etc., to achieve high sensitivity, good safety sexual effect

Inactive Publication Date: 2011-06-08
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, domestic HIV drug screening mainly uses infectious HIV virus to infect human lymphocyte cell lines. Since this cell model uses wild-type HIV virus, it is infectious to humans and must be strictly limited to negative pressure P3 laboratories. In addition, the system does not contain a reporter gene, resulting in complex detection procedures and low sensitivity

Method used

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  • HIV medicament screening cell model and special pseudotype lentivirus therefor
  • HIV medicament screening cell model and special pseudotype lentivirus therefor
  • HIV medicament screening cell model and special pseudotype lentivirus therefor

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0033] The drug screening of embodiment 1, HIV

[0034] 1. Construction of plasmid pLenti-Luc

[0035] see Figure 4The recombinant lentiviral plasmid pLenti-Luc containing the luciferase gene was constructed.

[0036] 1. Construction of intermediate plasmid pEGFP-N1-lac

[0037] Plasmid pUC19 was double-digested with BamHI and PvuII, and a 209bp fragment was recovered; PvuII and EcoRI double-digested plasmid pUC19, and a 92bp fragment was recovered; the above two fragments were mixed and ligated with the vector pEGFP-N1 that had been double-digested and dephosphorylated by BamHI and EcoRI , transform Escherichia coli DH5a, pick green colonies after IPTG induction, carry out enzyme digestion identification, obtain the recombinant plasmid pEGFP-N1-lac that can express EGFP protein in Escherichia coli and eukaryotic cells simultaneously (see Figure 5 ).

[0038] 2. Construction of intermediate plasmid pUC19-Luc

[0039] The luciferase gene-containing plasmid pG5Luc was dou...

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Abstract

The invention discloses an HIV drug screening cell model and special pseudotype slow virus thereof; the invention constructs recombinant plasmid for expressing Gag gene and Pol gene of HIV, recombinant slow-virus plasmid for expressing reporter gene and recombinant plasmid for expressing Rev gene of HIV; the three plasmids and recombinant plasmid for expressing glycoprotein gene of capsid G of herpetic stomatitis are transfected in the cells of mammals so as to obtain pseudotype slow virus; the pseudotype slow virus is used for infecting the cells of mammals, thus obtaining the HIV drug screening cell model based on the reporter gene. The HIV drug screening cell model provided by the invention uses the virus with one-time infection with good safety and the reporter gene leads the cell model to have super high sensitivity.

Description

technical field [0001] The invention relates to an HIV drug screening cell model and its special pseudotype lentivirus. Background technique [0002] Human immunodeficiency virus (human immunodeficiency virus, HIV), also known as HIV, the disease caused by HIV infection is called AIDS, and its full name is acquired immunodeficiency syndrome (acquired immunodeficiency syndrome, AIDS). Some or all of them are lost, the number of CD4+ cells decreases, and then opportunistic infections, tumors, etc. occur, and the clinical manifestations are various. The disease spreads quickly and has a high fatality rate, and it is currently incurable, which has attracted the attention of governments and societies in various countries. The prevalence of HIV is distributed worldwide. Africa is the birthplace and hardest hit area of ​​HIV, and Europe and America are also the main epidemic areas. In recent years, the prevalence of HIV in Asia has shown a trend of rapid growth. Since the first d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/63C12N15/867C12N7/01C12N5/10C12Q1/70C12Q1/02
Inventor 童贻刚李敬云张昕安小平周育森
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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