OBJECTIVETo identify whether GC-box (-348 to -338) in human insulin gene promoter is a key cis-acting element.

METHODSHuman insulin gene promoter was sub-cloned into secreted alkaline phosphatase (SEAP) reporter plasmid. The deletion and mutation of GC-box in insulin gene promoter was performed. The activity of human insulin gene promoter was determined by evaluating the activity of SEAP in the supernatant of cell culture after the reporter plasmids were transfected in beta cell line betaTC3.

RESULTDeletion and mutation of GC box in human insulin gene promoter did not result in significant changes of the activity of the promoter in betaTC3.

CONCLUSIONThe GC-box is not a key cis-acting element in human insulin gene promoter.

Alkaline Phosphatase ; genetics ; metabolism ; Base Sequence ; Cell Line ; Enhancer Elements, Genetic ; GC Rich Sequence ; Gene Expression Regulation ; Humans ; Insulin ; genetics ; Promoter Regions, Genetic ; Recombinant Fusion Proteins ; genetics ; metabolism ; Sequence Deletion ; Transcription, Genetic ; Transfection