DATA SHEET
Reagent: HIV-1 NL4-3 ΔEnv Vpr Luciferase Reporter Vector (pNL4-3.Luc.R-E-)
Catalog Number: 3418
Lot Number: 160163
Release C Category:
Provided: 5 μg of dried purified DNA stabilized in DNAstable PLUS
Cloning Vector: pUC18
Ampicillin resistant
Description: A HIV-1 NL4-3 luciferase reporter vector that contains defective Nef, Env and Vpr.
Special This construct is 16,396 bp including the insert. Characteristics: In this reporter vector, a firefly luciferase gene was inserted into the pNL4-3 nef gene while frameshifts in env and vpr render this clone Env and Vpr deficient.
To generate this plasmid, a frameshift near the 5'-end of env was introduced by using T4 DNA polymerase to fill in the NdeI site (nt 5950) of pNL4-3. This renders the clone Env deficient. A firefly luciferase gene was then inserted into the nef gene by removing the BamHI (nt 8021) to XhoI (nt 8443) fragment of pHXB-Luc (Chen et al. 1994) and ligating it to the same sites in env deficient pNL4-3. A frameshift was introduced in vpr by filling in the AflII site (nt 5180) corresponding to amino acid 26.
This construct is competent for a single round of replication and requires co-transfection with an Env expression vector to produce infectious virus.
Note: In this plasmid, Env was rendered deficient due to a small frameshift in the beginning of the Env gene. As a result, there is a possibility of recombinantion with another Env that could generate a viable virus. Please examine your sequences prior to generating pseudoviruses of any kind with this plasmid and exercise caution.
Contributor provided protocol and sequence information
ALL RECIPIENTS OF THIS MATERIAL MUST COMPLY WITH ALL APPLICABLE BIOLOGICAL, CHEMICAL, AND/OR RADIOCHEMICAL SAFETY STANDARDS INCLUDING SPECIAL PRACTICES, EQUIPMENT, FACILITIES, AND REGULATIONS. NOT FOR USE IN HUMANS.
REV: 05/21/2020 Page 1 of 2 Plasmid map and sequence file lot 160163
This reagent is currently being provided as dried purified DNA stabilized in DNAstable PLUS. Please see the notice for additional information and the protocol for reconstitution of dried DNA reagents. Dried DNA Notice
Plasmids can be propagated in STBL2 cells and grown at 37°C. Larger plasmids may benefit from growth at 30°C. This construct may also be grown in other competent cells.
Recommended Keep the reagent at room temperature in a dry storage cabinet or in a moisture barrier Storage: bag.
Contributor: Dr. Nathaniel Landau
References: Connor, R. I., Chen, B. K., Choe, S., & Landau, N. R. (1995). Vpr is required for efficient replication of human immunodeficiency virus type-1 in mononuclear phagocytes. Virology, 206(2), 935-944. doi: 10.1006/viro.1995.1016 PUBMED
He, J., Choe, S., Walker, R., Di Marzio, P., Morgan, D. O., & Landau, N. R. (1995). 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(11), 6705-6711. PUBMED
Chen, B. K., Saksela, K., Andino, R., & Baltimore, D. (1994). Distinct modes of human immunodeficiency virus type 1 proviral latency revealed by superinfection of nonproductively infected cell lines with recombinant luciferase-encoding viruses. J Virol, 68(2), 654-660. PUBMED
NOTE: Acknowledgment for publications should read “The following reagent was obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: HIV-1 pNL4-3 ΔEnv Vpr Luciferase Reporter Vector (pNL4-3.Luc.R-E-) from Dr. Nathaniel Landau.” Also include the references cited above in any publications. Patent pending. Requests from commercial organizations must be directed to the New York University Office of Industrial Liaison at the following email address: [email protected].
Last Updated May 21, 2020
ALL RECIPIENTS OF THIS MATERIAL MUST COMPLY WITH ALL APPLICABLE BIOLOGICAL, CHEMICAL, AND/OR RADIOCHEMICAL SAFETY STANDARDS INCLUDING SPECIAL PRACTICES, EQUIPMENT, FACILITIES, AND REGULATIONS. NOT FOR USE IN HUMANS.
REV: 05/21/2020 Page 2 of 2