Abstract
Swiss 3T3 fibroblasts were transfected with the provided plasmids of LacSwitch™ Inducible Mammalian Expression System (Stratagene). Stable transfectants were selected, expanded and characterised. At first, the production of CAT in these cell lines could be induced by IPTG treatment, but the inducibility was lost after a few months in culture in a reproducible manner. Further analysis revealed that the transfectants did not lose the cat gene nor the lac repressor protein. As a result, we conclude that LacSwitch™ Inducible Mammalian Expression System needs further modification for use in Swiss 3T3 fibroblasts.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Peles E, Nativ M, Campbell PL, Sakurai T, Martiniz R, Lev S, Clary DO, Schilling J, Barnea G, Plowman GD, Grumet M, Schlessinger J: The carbonic anhydrase domain of receptor tyrosine phosphatase beta is a functional ligand for the axonal cell recognition molecule contactin. Cell 82: 251–260, 1995
Brown-Shimer S, Johnson DA, Hill DE, Bruskin AM: Effect of protein tyrosine phosphatase 1B expression on transformation by the human neu oncogene. Cancer Res 52: 478–482, 1992
Celler JW, Luo X, Gonez LJ, Böhmer FD: mRNA expression of two transmembrane protein tyrosine phosphatases is modulated by growth factors and growth arrest in 3T3 fibroblasts. Biochem Biophys Res Commun 209: 614–621, 1995
Stratagene, LacSwitchTM Inducible Mammalian Expression System, instruction manual, Catalog #217450, 1993
Matthews RJ, Cahir ED, Thomas ML: Identification of an additional member of the protein-tyrosine-phosphatase family: Evidence for alternative splicing in the tyrosine phosphatase domain. Proc Natl Acad Sci USA 87: 4444–4448, 1990
Brown M, Figge J, Hansen U, Wright C, Jeang K, Khoury G, Livingston DM, Roberts TM: lac repressor can regulate expression from a hybrid SV40 early promoter containing a lac operator in animal cells. Cell 49: 603–612, 1987
Miller JH, Reznikoff WS: The Operon. Cold Spring Harbor, New York, 1980
Figge J, Wright C, Collins CJ, Roberts TM, Livingston DM: Stringent regulation of stably integrated chloramphenicol acetyl transferase genes by E. coli lac repressor in monkey cells. Cell 52: 713–722, 1988
Barkley MD, Bourgeois S: In: J.H. Miller, W.S. Reznikoff (eds). The Operon. Cold Spring Harbor, New York, 1980, pp 177–220
Wyborski DL, Short JM: Analysis of inducers of the E. coli lac repressor system in mammalian cells and whole animals. Nucl Acid Res 19: 4647–4653, 1991
Yarranton GT: Inducible vectors for expression in mammalian cells. Curr Opin Biotech 3: 506–511, 1992
Auckland-Berglund CE, Leib DA: Efficacy of tetracycline-controlled gene expression is influenced by cell type. BioTechniques 18: 196–200, 1995
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Luo, X., Celler, J.W. & Berndt, A. LacSwitchTM Inducible Mammalian Expression System in mouse Swiss 3T3 fibroblasts. Mol Cell Biochem 200, 127–132 (1999). https://doi.org/10.1023/A:1007079913318
Issue Date:
DOI: https://doi.org/10.1023/A:1007079913318