PA317 cyclin E-S (ATCC® CRL-2188)

Organism: Mus musculus, mouse  /  Cell Type: fibroblast  /  Disease: sarcoma

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Organism Mus musculus, mouse
Cell Type fibroblast
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Disease sarcoma
Age embryo
Strain NIH/Swiss
Applications
The LXSN vector uses the 5'long terminal repeat of Moloney murine sarcoma virus to express the inserted cDNA and contains the neomycin phosphotransferase gene (G418 resistance) as a selectable marker.
The PA317 cyclin E-S cell line expresses human cyclin E-S.
Fibroblasts engineered to constitutively overexpress either form of cyclin E show elevated cyclin E dependent kinase activity and a shortened G1 phase of the cell cycle.
Derivation
PA317 cyclin E-S is an amphotropic retrovirus producing cell line constructed from the PA317 (see ATCC CRL-9078) fibroblast cell line by introduction of a retrovirus expression vector (LXSN).
Genes Expressed
cyclin E-S
Comments
PA317 cyclin E-S is an amphotropic retrovirus producing cell line constructed from the PA317 (see ATCC CRL-9078) fibroblast cell line by introduction of a retrovirus expression vector (LXSN).
The LXSN vector uses the 5'long terminal repeat of Moloney murine sarcoma virus to express the inserted cDNA and contains the neomycin phosphotransferase gene (G418 resistance) as a selectable marker.
The full length cDNA coding human cyclin E-S (1.6 kb HindIII fragment) was treated with Klenow fragment and cloned into the HpaI site of the vector.
The PA317 cyclin E-S cell line expresses human cyclin E-S.
Cyclin E is a nuclear protein essential for the G1 to S phase transition.
Mammalian cells express two forms of cyclin E protein which differ from each other by the presence (cyclin E-L) or absence (cyclin E-S) of a 15 amino acid amino terminal domain.
Fibroblasts engineered to constitutively overexpress either form of cyclin E show elevated cyclin E dependent kinase activity and a shortened G1 phase of the cell cycle.
Complete Growth Medium Dulbecco's modified Eagle's medium with 4.5 g/L glucose containing 0.1 mg/ml G-418, 90%; calf serum, 10%
Subculturing
Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:10 is recommended
Medium Renewal: Twice per week
Remove medium, rinse flask with fresh 0.25% trypsin, 0.02% EDTA and allow the flask to sit at room temperature until the cells detach. Add fresh medium, aspirate and dispense into new flasks.
Cryopreservation
Culture medium, 95%; DMSO, 5%
Name of Depositor JM Roberts
References

Ohtsubo M, et al. Human cyclin E, a nuclear protein essential for the G1-to-S phase transition. Mol. Cell. Biol. 15: 2612-2624, 1995. PubMed: 7739542

Ohtsubo M, Roberts JM. Cyclin-dependent regulation of G1 in mammalian fibroblasts. Science 259: 1908-1912, 1993. PubMed: 8384376

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
References

Ohtsubo M, et al. Human cyclin E, a nuclear protein essential for the G1-to-S phase transition. Mol. Cell. Biol. 15: 2612-2624, 1995. PubMed: 7739542

Ohtsubo M, Roberts JM. Cyclin-dependent regulation of G1 in mammalian fibroblasts. Science 259: 1908-1912, 1993. PubMed: 8384376