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微信小章| 产品名称： | HIIF-D R10C101-250nM |
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| 商品货号： | TS212005 |
| Organism： | Cricetulus griseus, hamster, Chinese |
| Tissue： | ovary |
| Product Format： | frozen |
| 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. |
| Gender： | female |
| Applications： | in another host, produces protein interferon gamma |
| Disclosure： | This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC. |
| Derivation： | The HIIF-D line was established from CHO DHFR- cells (deficient in dihydrofolate reductase) co-transformed with pSV-IFN-gamma (containing the coding sequence for human interferon gamma) and pAdD26SV(A)-3 (containing the mouse DHFR coding sequences).
Constructed by replacing the HindIII/BglII fragment containing murine Dfhr with the IFNG sequence.
The pBR322-derived sequences were also modified by replacing a EcoRI/PvuII fragment that inhibits replication in mammalian cells with an EcoRI/SalI fragment from pML.The pBR322-derived sequences were also modified by replacing a EcoRI/PvuII fragment that inhibits replication in mammalian cells with an EcoRI/SalI fragment from pML.
Also contains pAdD26SV(A)-3 which was cotransformed to express murine Dhfr. Both plasmids were digested with EcoRI before transformation. This clone was selected after growth in 250 nM methotrexate. |
| Clinical Data： | female |
| Genes Expressed： | human interferon gamma |
| Cellular Products： | human interferon gamma |
| Comments： | The cells produce human gamma interferon.
The order of the major features in this plasmid is: SalI - SV40 promoter - HindIII - IFNG - SV40t - SV40 polyadenylation - EcoRI - ampR - ori.
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| Complete Growth Medium： | Alpha minimum essential medium without nucleosides and with 250 nM methotrexate; 90%; fetal bovine serum, 10%
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| Subculturing： | Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
xa0xa0xa0xa0xa0xa0xa0 Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach.xa0 Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
Medium Renewal: 2 to 3 times per week |
| Cryopreservation： | Freeze medium: complete growth medium, 95%; DMSO, 5% |
| Culture Conditions： | Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
| Name of Depositor： | Biogen, Inc., Biogen N.V. |
| U.S. Patent Number： | |
| References： | Fiers WC, Allet B. DNA sequences, recombinant DNA molecules and processes for producing human gamma interferon-like polypeptides in high yields. US Patent 5,004,689 dated Apr 2 1991 |