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SPECIES: mouse, NIH Swiss
ECACC catalog code: 95050229
Transfected by: Transfection Back To Transfection WebPage
Description: Mouse NIH Swiss embryo, transfected with retroviral DNA
Viruses: See Properties
Depositor: Dr Y Takahara, Central Research Laboratories, Ajinomoto Co., Kawasaki, JAPAN (Originator)
No restrictions. Patent: None Specified By Depositor
Properties: Applications: Retrovirus packaging, gene transfer assays
Available in the following LABORATORY:
Part of the Business Division of the Health Protection Agency (ECACC, Salisbury, Wiltshire)
DMEM + 2mM Glutamine + 250µg/ml G418 + 10% Fetal Bovine Serum (FBS). Split confluent cultures 1:2 to 1:6 i.e. seeding at 2-5x10,000 cells/cm2 using 0.25% trypsin or trypsin/EDTA; 5% CO2; 37C.
The retrovirus packaging Ampli-GPE (clone A57) cells were obtained from NIH3T3 fibroblasts (ECACC No. 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virus were separated onto two different plasmids in which the 5' long terminal repeat (LTR) and the 3' LTR had been replaced by the mouse metallothionein I promoter or the rabbit ß-globin gene.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.
Research council deposit: No;
Price Code: C;
Validated by: isoenzymes (species confirmation)
- J Virol 1992;66:3725 - PMID: 1316479
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Cell Line Data Base: structure and recent improvements towards molecular authentication of human cell lines.
Nucleic Acids Research 2009 37(Database issue):D925-D932.