Cloning of the Promoter Regions of Golgi Membrane Glycoprotein GP73
摘要
[Objective] The aim of this study is to clone the active promoter of golgi membrane glycoprotein GP73. [Method] The sequence within the range of upstream 1 000 bp and downstream 400 bp of transcription initiation site was analyzed, the genomic DNA of hepatoma cell line Huh-7 was regarded as template for PCR amplification. The amplified fragment was cloned into recombinant construct with enhanced green fluorescent protein (EGFP) report gene. The expression of EGFP in recombinants were observed under fluorescence microscope after transfection, with the assistant of flow cytometry. [Result] The 1 310 bp ranging between upstream 980 bp and downstream 330 bp of transcription initiation site assumed promoter function. The region contains two core promoters and several conserved sequences including TATA box and many DNA binding sites such as NF-κB, AP1, GC-SP1. [Conclusion] The cloning of the promoter provides a reference for the study of the transcription mechanism of GP73.