Patent ID: 6465246
Filing Date: 2002-10-15
Classification: A61K,C07K,C12N

Abstract:
A nucleic acid construct for expressing an effector protein, comprising:(a) a first promoter, wherein said first promoter comprises the promoter of the cdc25C gene (nucleic acids âˆ’290 to +121); (b) a polynucleotide encoding a transcription factor, wherein the expression of the transcription factor is controlled by the first promoter, and wherein said transcription factor comprises the nuclear localization signal (NLS) of SV40 (SV40 large T, amino acids 126 to 132, PKKKRKV (SEQ ID NO: 13), the acid transactivation domain (TAD) of HSV-1 VP16 (amino acids 406 to 488), the RB binding sequence of the E2F-1 protein (amino acids 409 to 426 (LDYHFGLEEGEGIRDLFD) (SEQ ID NO: 14), and the DNA binding domain of the Gal4 protein (amino acids 1 to 147); (c) a second promoter, to which the transcription factor binds, wherein said second promoter comprises 10Ã— the binding sequence for the Gal4 DNA-binding sequence having the nucleotide sequence 5â€²-CGGACAATGTTGACCG-3â€² (SEQ ID NO: 1) and the SV40 basal promoter (nucleic acids 48 to 5191); and (d) a polynucleotide encoding the effector protein, comprising the sequence GCCACC (SEQ ID NO: 17), the cDNA for the immunoglobulin signal peptide (nucleotide sequence 63 to 107) and the cDNA for &THgr;-glucuronidase (nucleotide sequence 93 to 1982), wherein the expression of the effector protein is controlled by the second promoter, wherein the activity of the transcription factor depends on one or more cellular regulatory proteins that bind to the transcription factor and affect the activity thereof.