Merck, 95050229-DNA-5UG, Ampli-GPE

Application
Retrovirus packaging, gene transfer assays
Biochem/physiol Actions
Mouse NIH Swiss embryo, transfected with retroviral DNA
The retrovirus packaging Ampli-GPE (clone A57) cells were obtained from NIH3T3 fibroblasts (Sigma Catalogue number. 93061524) being co-transfected with two kinds of recombinant chimeric proviral genes in the BMG Neo vector. To reduce the homology between proviral DNA in the packaging cell and the retroviral vector, the gag-pol and env genes of Moloney murine leukaemia virus were separated onto two different plasmids in which the 5′ long terminal repeat (LTR) and the 3′ LTR had been replaced by the mouse metallothionein I promoter or the rabbit β-globin gene. This expression and amplification system produces a viral titre high enough for gene transfer to somatic cells (up to 5x1,000,000 CFU/ml) and avoids the generation of replication-competent viruses.
Preparation Note
DMEM + 2mM Glutamine + 250μg/ml G418 + 10% Foetal Bovine Serum (FBS).
Split sub-confluent cultures (70-80%) 1:2 to 1:6 i.e. seeding at 2-5x10,000 cells/cm² using 0.25% trypsin or trypsin/EDTA; 5% CO₂; 37°C.
Other Notes
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This cell line is part of the European Collection of Authenticated Cell Cultures (ECACC), an international repository managed by the United Kingdom Health Security Agency (UKHSA). No licensing agreement is required when either this cell line or the DNA extracted from it are used for internal research purposes only. Any other use of these products is prohibited without the express written permission of UKHSA. Inquiries regarding authorized use of this cell line or its genomic DNA may be directed to [email protected].