Aim To develop a method of
drug screening ba sed on reporter gene and the signal transduction of interferon-α system in
order to scre en the small molecular compounds with Interferon-α-like activity.Meth ods A recombinant vector pTAL/ISRE-SEAP was constructed by inserting a synthetic sequence composed of five IFN-α bounded elements in front of promot er of pTAL/SEAP vector. pTAL/ISRE-SEAP was then
transfected into ECV304 cells. HygromycinB(500 mg·L -1) was added 48 h after transfection to select positive clones that can be induced by IFN-α to express reporter gene SEAP. Stably tra nsfected cell clones were isolated and used to screen compound banks for the com pounds with Interferon-α-like activity.The speciality was tested by the cytokines of same family (IFN-β;IFN-γ) and growth factor (eg parathyroid factor). Results Stably transfected clones were obtained.The expression of reporter gene SEAP from one of the cloned cells was induced by IFN-α in do se-dependent manner.The expression level of induction by IFN-β,IFN-γ and PT H was low, indicating the high speciality of the developed method.The Z'-facto r(0.8) showed that the method is very stable .Conclusion Stabl y transfected positive clone cells line transfected with recombinant vector pTAL /ISRE-SEAP were obtained.The expression of SEAP was specially induced by IFN- α The cloned cells can be used to screen for compounds with interferon-α-lik e activity by testing luminescent value of expressed SEAP in wells of microlitr e plate.