Lac/Tet dual-inducible system functions in mammalian cell lines

H. S. Liu, C. H. Lee, C. F. Lee, I. J. Su, T. Y. Chang

研究成果: Article同行評審

17 引文 斯高帕斯(Scopus)

摘要

The Escherichia coli Lac repressor (Lac system) and tetracycline responsive promoter (Tet system) systems have been used individually to regulate gene expression at the cellular as well as at the organismal levels. In this study, these two systems were combined (designated Lac/Tet dual- inducible system) to regulate two inducible genes simultaneously in a single cell. The isopropyl-β-D-thiogalactopyranoside (IPTG) and tetracycline (used for the operation of the Lac and the Tet systems) were non-cytotoxic to the cells when added together into the cells at around the optimal concentrations (IPTG: ≤5 mM; tetracycline: <1.5 μg). The rate and efficiency of induction and repression of two inducible genes regulated by the Lac/Tet dual-inducible system were similar to the results obtained when one inducible gene is regulated by one inducible system in a single cell. The Lac-Tet dual- inducible system could function in many cell lines, which was demonstrated by regulating the expression of β-galactosidase and luciferase reporter genes in five tumor cell lines by transient transfection analysis. The feasibility of introducing a second inducible system into an already established inducible cell line was confirmed. Finally, we showed that the Lac/Tet dual- inducible system functions at translational and at functional levels in a stable cell line named 7-4-b, which contains the Ha-ras and bcl-2 inducible genes. In conclusion, this study extends the application of prokaryotic inducible systems from the regulation of a single gene to two genes and helps clarify the relationship between two genes and the effects of two genes on the cells.

原文English
頁(從 - 到)624-632
頁數9
期刊BioTechniques
24
發行號4
DOIs
出版狀態Published - 1998

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

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