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Room temperature, protected from moisture
Ψ -Moloney Murine Leukemia virus (MLV) DNA (from Richard Mann) was cloned into the SV40 expression vector pSV7d at the EcoRI site. Restriction sites in the MLV sequence can be found in the Appendix to Volume 2 of the Cold Spring Harbor Tumor Virus Book. The mouse flanking sequences present on either side of the provirus have not been sequenced.
This construct is 13,966 base pairs including the insert; the insert is 11,606 base pairs. The cloning vector used to create pSV-Ψ-MLV-env-was pSV7d and the resulting vector is ampicillin resistant. For additional details please refer to the provided vector map and sequence files.
This plasmid is used to produce MLV retroviral vectors using the method of Landau and Littman. In the original method, COS cells were transfected with this plasmid and an amphotropic MLV env vector. Higher virus titers can be obtained by transfecting 293 cells instead of COS cells, and with the VSV-G expression vector instead of A-MLV.
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.
Each vial of ARP-3422 contains approximately 5 µg of dried purified DNA stabilized in DNAstable®PLUS. Please refer to the appropriate data sheet for lot-specific information.
Landau, N. R. and D. R. Littman. “Packaging System For Rapid Production Of Murine Leukemia Virus Vectors With Variable Tropism.” J. Virol. 66 (1992): 5110-5113. PubMed:1321291.
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