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HIV medicament screening cell model and special pseudotype lentivirus therefor

A technology of lentivirus and recombinant lentivirus, which is applied to cells modified by introducing foreign genetic material, viruses/phages, applications, etc. safety effect

Inactive Publication Date: 2009-01-28
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

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Embodiment 1

[0033] Example 1. Drug screening for 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 double-di...

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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 present invention relates to a HIV drug screening cell model and its special pseudotyped lentivirus. Background technique [0002] Human immunodeficiency virus (HIV), also known as HIV, the disease caused by HIV infection is called AIDS, the full name of acquired immunodeficiency syndrome (AIDS), the immune function of patients with this disease Partial or complete loss, the number of CD4+ cells is reduced, and then opportunistic infections, tumors, etc., have various clinical manifestations. The disease spreads rapidly, has a high fatality rate, and is currently incurable, which has attracted the attention of governments and societies around the world. The prevalence of HIV is distributed worldwide. Africa is the birthplace and hardest hit area of ​​HIV, and Europe and the Americas are also major endemic areas. In recent years, the prevalence of HIV in Asia has shown a trend of rapid growth. Since the first discovery of HIV-infected people in my...

Claims

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Application Information

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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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