Improved ecdysone receptor-based inducible gene regulation system

Subba R. Palli, Mariana Z. Kapitskaya, Mohan B. Kumar, Dean E. Cress

Research output: Contribution to journalArticlepeer-review

81 Scopus citations


To develop an ecdysone receptor (EcR)-based inducible gene regulation system, several constructs were prepared by fusing DEF domains of Choristoneura fumiferana EcR (CfEcR), C. fumiferana ultraspiracle (CfUSP), Mus musculus retinoid X receptor (MmRXR) to either GAL4 DNA binding domain (DBD) or VP16 activation domain. These constructs were tested in mammalian cells to evaluate their ability to transactivate luciferase gene placed under the control of GAL4 response elements and synthetic TATAA promoter. A two-hybrid format switch, where GAL4 DBD was fused to CfEcR (DEF) and VP16 AD was fused to MmRXR (EF) was found to be the best combination. It had the lowest background levels of reporter gene activity in the absence of a ligand and the highest level of reporter gene activity in the presence of a ligand. Both induction and turn-off responses were fast. A 16-fold induction was observed within 3 h of ligand addition and increased to 8942-fold by 48 h after the addition of ligand. Withdrawal of the ligand resulted in 50% and 80% reduction in reporter gene activity by 12 h and 24 h, respectively.

Original languageEnglish
Pages (from-to)1308-1315
Number of pages8
JournalEuropean Journal of Biochemistry
Issue number6
StatePublished - Mar 2003


  • EcR
  • Gene switch
  • Ponasterone A
  • RXR
  • Receptors

ASJC Scopus subject areas

  • Biochemistry


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