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Quantity limit per order for this item is 1. This item can be ordered twice a year. Orders over this limit will be sent to NIAID for approval before shipment.
ARP-1067 (HIV-gpt) contains intact HIV-1HXB2 rev (a transactivating protein that is essential to the regulation of HIV-1 protein expression) and tat (trans acting activator of transcription), genes. Deletion of sequences encoding gp160 has rendered HIV-gpt replication-defective. The HIV-gpt construct is 14,294 base pairs and contains the SV40 origin of replication and coding sequences for the gpt gene in the the PvuII-DraI SV2gpt fragment. An XbaI-HpaI pHXB2gpt fragment (Drs. A. Fisher and F. Wong-Staal) containing proviral and flanking cellular sequences was cloned into the HincII-XbaI site of Bluescript pBS KS +/-. A 1.2 Kb NdeI-BglII fragment (nt 6402-7620) was deleted from the env gene, and the 1.1 kb PvuII-DraI SV2gpt fragment (Dr. M. Mulligan) was inserted at the env deletion site.
ARP-1067 contains the intact HXB2 rev and tat genes and is replication-defective. By itself, HIV-gpt produces non-infectious HIV-1 particles. Co-transfection of HIV-gpt with an envelope expression vector into COS cells results in the packaging of the replication-defective genome into infectious virions (virus is transiently produced). HIV or other retroviral env genes can be used to complement HIV-gpt to yield virus with the host range of the complementing gene. The gpt gene provides a convenient selection marker, since each successful infection leads to the growth of a drug-resistant (mycophenolic acid) colony.
Escherichia coli, strain HB101 is the recommended host for this vector, however, other bacterial strains should also be successful.
ARP-1067 was produced using the following cloning strategy: An XbaI - HpaI fragment from pHXB2gpt containing HIV-1 proviral and flanking cellular sequences was cloned into the HincII - XbaI site of pBS. A 1.2 kb NdeI - BglII fragment (nt 6402-7620) was deleted from the env gene, and the 1.1 kb PvuII - DraI SV2gpt fragment was inserted at the env deletion site. The pro virus, HIV-1 plasmid pHXB2gpt, was obtained from Dr. A. Fisher and Dr. F. Wong-Staal and pSV2gpt was obtained from Dr. M. Mulligan. Please see the plasmid map and sequence file for additional details.
Each vial of ARP-1067 contains approximately 5 µg of dried purified DNA stabilized in DNAstable®PLUS. Please refer to the appropriate data sheet for lot-specific information.
Page, K. A., N. R. Landau and D. R. Littman. “Construction and Use of a Human Immunodeficiency Virus Vector for Analysis of Virus Infectivity.” J. Virol. 64 (1990): 5270-5276. PubMed:2214018.
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