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Room temperature, protected from moisture
ARP-3419 [human immunodeficiency virus 1 (HIV-1) NL4-3 heat stable antigen reporter vector (pNL4-3.HSA.R+)] encodes replication competent NL4-3 proviral DNA.
Murine heat stable antigen CD24 (HSA) gene was inserted into the NotI/XhoI cloning site of pNL4-3 (BEI Resources ARP-114) nef gene to produce this clone. This construct is 14,971 base pairs including the insert which is 236 base pairs. pNL4-3.HSA.R+ is ampicillin resistant. The GenBank accession number for pNL4-3 is M19921. For additional details please refer to the provided vector maps and sequence files.
Virus can be produced by transfecting 2 × 106 293 or 293T cells with 20 µg NL4-3 DNA. Transfections can be performed in a 10 cm2 tissue culture dish using standard calcium phosphate protocols. Virus is typically harvested 48 hours post-transfection. Infections should be performed in a total volume of 0.5 mL. The amphotropic pseudotypes generally have much higher infectivity than those bearing HIV-1 env. Cultures infected with HSA viruses can be assayed by FACS analysis with a commercial CD24 antibody (Pharmingen) 2-5 days post-infection.
Each vial of ARP-3419 contains approximately 5 µg of dried purified DNA stabilized in DNAstable®PLUS. Please refer to the appropriate data sheet for lot-specific information.
He, J., et al. “Human Immunodeficiency Virus Type 1 Viral Protein R (Vpr) Arrests Cells in The G2 Phase of the Cell Cycle by Inhibiting P34cdc2 Activity.” J Virol. 69 (1995): 6705-6711. PubMed: 7474080.
Connor, R. I., et al. “Vpr is Required for Efficient Replication of Human Immunodeficiency Virus Type-1 in Mononuclear Phagocytes.” Virology 206 (1995):935-944. PubMed: 7531918.
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