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| 产品名称： | PA317 cyclin E-S |
|---|---|
| 商品货号： | TS211141 |
| Organism： | Mus musculus, mouse |
| Tissue： | embryo |
| Cell Type： | fibroblast |
| Product Format： | frozen |
| Morphology： | fibroblast |
| Culture Properties： | adherent |
| Biosafety Level： | 1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Disease： | sarcoma |
| Age： | embryo |
| Strain： | NIH/Swiss |
| Applications： | The LXSN vector uses the 5long 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 |
| Cellular Products： | 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 5long 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： | Dulbeccos modified Eagles medium with 4.5 g/L glucose containing 0.1 mg/ml G-418, 90%; calf serum, 10%
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| 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 |
| Deposited As： | Mus musculus |
| 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 |