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ECACC catalog code: 95051229
Description: Human (HeLa) with tTA
Viruses: See Properties
Depositor: Dr M Gossen and Prof Dr H Bujard, Zentrum für Moleculaire Biologie, University Heidelberg, GERMANY (Originator)
No restrictions. Patent: None Specified By Depositor
Properties: Applications: Specific and stringent control of the activity of an individual gene (luciferase) in a mammalian cell can be monitored. It is also suitable for creation of 'on/off' situations and analysis of mRNA decay rates under physiological conditions.
Available in the following LABORATORY:
Part of the Business Division of the Health Protection Agency (ECACC, Salisbury, Wiltshire)
EMEM (EBSS) + 2mM Glutamine + 1% Non Essential Amino Acids (NEAA) + 1% Vitamins + 500µg/ml G418 + 300µg/ml Hygromycin + 10% Fetal Bovine Serum (FBS). Freeze cells in 40% medium, 10% Dimethyl Sulphoxide (DMSO) and 50% Fetal Bovine Serum (FBS). Split confluent cultures 1:5 to 1:20 i.e. seeding at 5x1,000 to 2x10,000 cells/cm2 using 0.25% trypsin or trypsin/EDTA. Alternatively cells can be washed with PBS and subcultured after addition of PBS with 0.8mM EDTA (30% of original volume) ; 5% CO2; 37C.
X1/5 is a HeLa cell line stably transfected with a 'tet' repressor fused with the activating domain of virion protein 16 of herpes simplex virus thus generating a tetracycline-controlled transactivator (tTA). This transactivator stimulates transcription from a minimal promoter sequence derived from the human cytomegalovirus promoter IE combined with 'tet' operator sequences. Integration of a luciferase gene controlled by a tTA - dependent promoter into this tTA - producing HeLa cell line leads to high levels of tetracycline-sensitive luciferase expression. The luciferase activity can be regulated over up to five orders of magnitude depending on the concentration of tetracycline in the culture medium (0-1µg/ml).
Research council deposit: No;
Price Code: C;
Validated by: isoenzymes (species confirmation)
- Proc Natl Acad Sci U S A 1992;89:5547
For information, get in touch with:
IRCCS Ospedale Policlinico San Martino
Largo Rosanna Benzi, 10 - I-16132 Genova - Italy
Dott.ssa Barbara Parodi
If you want to cite CLDB and HyperCLDB, please use the following reference:
P. Romano, A. Manniello, O. Aresu, M. Armento, M. Cesaro, B. Parodi.
Cell Line Data Base: structure and recent improvements towards molecular authentication of human cell lines.
Nucleic Acids Research 2009 37(Database issue):D925-D932.