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微信小章| 产品名称： | CHO-1C6 |
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| 商品货号： | TS212360 |
| Organism： | Cricetulus griseus, hamster, Chinese |
| Tissue： | ovary |
| Product Format： | frozen |
| Morphology： | epithelial |
| 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. |
| Applications： | produces protein glucose-6-phosphate isomerase neuroleukin |
| Derivation： | This cell line was produced by cotransfection with neuroleukin and dihydrofolate reductase transcription units. pAdD26SVp(A)3 was constructed by cloning the mouse dihydrofolate reductase cDNA from the plasmid pAdD26-1 into the vector, pSVOd. A 900 bp BclI/PstI fragment of SV40 containing the early polyadenylation site was inserted. |
| Genes Expressed： | glucose-6-phosphate isomerase(neuroleukin) Nlk*,glucose-6-phosphate isomerase |
| Comments： | Cotransfection and cointegration of pAdNLK and pAdD26SVp(A)3 enables the SV40 enhancer element of pAdNLK to activate the mouse dihydrofolate reductase transcription unit of pAdD26SVp(A)3. This cell line was produced by cotransfection with neuroleukin and dihydrofolate reductase transcription units. Production of neuroleukin was increased by selection with methotrexate. Recently, it has been shown that the polypeptide neuroleukin is in fact glucophosphoisomerase. pAdNLK contains the SV40 enhancer, the mouse neuroleukin cDNA, the mouse dihydrofolate reductase cDNA, the SV40 polyadenylation sequence, and pBR322 sequences minus the inhibitory sequences from 1424 to 2490. pAdD26SVp(A)3 contains the adenovirus major late promoter (AdMLP), the mouse dihydrofolate reductase cDNA, the SV40 polyadenylation sequence, pBR322 sequences minus the inhibitory sequences from 1424 to 2490, and the SV40 origin of replication. The pAdNLK insert contains the complete nucleotide sequence and 5 and 3 untranslated sequences. The cell line CHO-IC6 also contains pAdD26SVp(A)3. Selection of the parental line for growth in methotrexate (MTX) coamplified MTX resistance and neuroleukin secretion. CHO-IC6 secretes 4.4 ug neuroleukin per 10(7) cells per 24 hr. pAdD26SVp(A)3 was constructed by cloning the mouse dihydrofolate reductase cDNA from the plasmid pAdD26-1 into the vector, pSVOd. A 900 bp BclI/PstI fragment of SV40 containing the early polyadenylation site was inserted. |
| Complete Growth Medium： | Alpha minimum essential medium with 2 mM L-glutamine and 20 nM methotrexate, 90%; fetal bovine serum, 10%
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| Subculturing： | Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended Medium Renewal: 2 to 3 times per week Remove medium, rinse monolayer with fresh 0.25% trypsin solution, remove trypsin and let the culture sit at room temperature (or 37C) until the cells detach (about 10 minutes). Add fresh medium, aspirate and dispense into new flasks. |
| Name of Depositor： | M Gurney |
| References： | Kaufman RJ, Sharp PA. Construction of a modular dihydrofolate reductase cDNA gene: analysis of signals utilized for efficient expression. Mol. Cell. Biol. 2: 1304-1319, 1982. PubMed: 6131378 Kaufman RJ, et al. Coamplification and coexpression of human tissue-type plasminogen activator and murine dihydrofolate reductase sequences in Chinese hamster ovary cells. Mol. Cell. Biol. 5: 1750-1759, 1985. PubMed: 4040603 Gurney ME, et al. Molecular cloning and expression of neuroleukin, a neurotrophic factor for spinal and sensory neurons. Science 234: 566-574, 1986. PubMed: 3764429 Itagaki E, et al. Neuroleukin: a lymphokine product of lectin-stimulated T cells. Science 234: 574-581, 1986. PubMed: 3020690 Chaput M, et al. The neurotrophic factor neuroleukin is 90% homologous with phosphohexose isomerase. Nature 332: 454-455, 1988. PubMed: 3352744 Mark Gurney, personal communication |