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微信小章| 产品名称： | MB19tsA, clone 2B2 Mbeta19tsA, clone 2B2 |
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| 商品货号： | TS211518 |
| Organism： | Mus musculus, mouse |
| Tissue： | embryo |
| Cell Type： | fibroblast SV40 large T antigen transfected |
| Product Format： | frozen |
| Morphology： | fibroblast |
| Culture Properties： | adherent |
| Biosafety Level： | 2 xa0Cells contain SV-40 viral sequences
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. |
| Age： | 10.5 days gestation |
| Applications： | The cell line is deficient in base excision repair due to homozygous deletion of the promoter and exon I in the pol-bet a gene. This deletion event is reversible and therefore this cell line is ideal for site-specific recombination using the Cre-lox system. |
| Storage Conditions： | liquid nitrogen vapor phase |
| Derivation： | MB19tsA, clone 2B2 is a fibroblast-like cell line derived from primary cultures of DNA polymerase beta (pol-beta) deficient fibroblasts isolated from embryos from heterozygous females ( beta-polymerase +/-) at day 10.5 of gestation. |
| Comments： | The genotype of this cell line has recently been determined to be DNA polymerase beta mutant (null) and DNA polymerase iota mutant (null).
The pol-beta deficient fibroblasts were immortalized by transfection with the DNA plasmid construct ptsA58H which contains coding sequences for both a hygromycin-resistance gene and tsA58H, a temperature-sensitive SV40 T antigen.
Cells were selected in the presence of 0.080 mg/ml hygromycin for 21 days and further selected by limiting dilution.
The cell line is deficient in base excision repair due to homozygous deletion of the promoter and exon I in the pol-bet a gene.
The deletion of pol-beta is mediated by the Cre-loxP recombination.
This deletion event is reversible and therefore this cell line is ideal for site-specific recombination using the Cre-lox system.
The contributor of this line has demonstrated that insertion of a single-copy of a variety of cDNAs can be site-specifically reinserted at the pol-beta locus, and that this is a stable event. |
| Complete Growth Medium： | Dulbeccos modified Eagles medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and supplemented with 0.08 mg/ml hygromycin B, 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.
Subcultivation Ratio: 1:10 to 1:15 Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994. |
| Cryopreservation： | Complete growth medium described above supplemented with 5% (v/v) DMSO.xa0 Cell culture tested DMSO is available as ATCC Catalog No. 4-X. |
| Culture Conditions： | Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5% |
| Name of Depositor： | RW Sobol, SH Wilson |
| References： | Gu H, et al. Deletion of a DNA polymerase beta gene segment in T cells using cell type-specific gene targeting. Science 265: 103-106, 1994. PubMed: 8016642 Sobol RW, et al. Requirement of mammalian DNA polymerase-beta in base-excision repair. Nature 379: 183-186, 1996. PubMed: 8538772 Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC. Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988. Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC. Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online. |