31] Use of tetracycline operator system to regulate oncogene expression in mammalian cells

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This chapter describes the use of tetracycline operator system to regulate oncogene expression in mammalian cells. Two inducible prokaryotic systems that have recently received attention are the Lac activator protein and the use of the operator of the tetracycline resistance genes. The chapter focuses on the use of the operator of the tetracycline resistance genes. In Escherichia coli, the tetracycline repressor (tetR) binds the tetracycline operator tetO to prevent transcription of tetracycline resistance genes in the absence of the drug tetracycline. The tetR has high specificity for the tetO, whereas tetracycline has high affinity for the tetR. In the absence of tetracycline, the tetR binds the tetO, effectively blocking transcription by steric interference. The binding of tetracycline by the tetR, however, abrogates this transcriptional repression as the consequent conformational change in the DNA binding domain of the protein releases the tetR from its association with the tetO. A tetracycline-transactivating system based on the tetR has been developed and is described in the chapter.