Constitutive expression of human coagulating factor IX in HeLa cells by homologous recombination of the promoter

作     者：杨晓青 王飞 王颖 卢大儒 邱信芳 薛京伦 

作者机构：1. State Key Laboratory of Genetics Engineering Institute of Genetics School of Life Sciences Fudan University 200433 Shanghai China 

出 版 物：《Science China(Life Sciences)》 (中国科学（生命科学英文版）)

年 卷 期：2001年第44卷第1期

页      面：18-24页

核心收录：

学科分类：0710[理学-生物学] 07[理学] 08[工学] 09[农学] 071007[理学-遗传学] 0901[农学-作物学] 0836[工学-生物工程] 090102[农学-作物遗传育种] 

基　　金：the State High Technology Program "863"  the National Natural Science Foundation of China (Grant No. 39880019) and Shanghai Scientific Phosphor Program 

主　　题：human clotting factor IX constitutive expression homologous recombination promoter. 

摘      要：Constitutive expression of hFIX protein in nonhepatocytes wasstudied. The gene targeting vector was constructed and transferred into HeLa cells. With the detection system of PCR, we demonstrated that the endogenous hFIX promoter was replaced with an hCMV promoter when targeted insertion of the constructor was directed by the sequence homology. The expression of hFIX in the modified HeLa cells, 11.2 ng/106 cell/24 h, strongly suggested that hFIX gene could be activated by a powerful promoter in nonhepatocytes. The results would make it possible to examine the feasibility of re-regulate gene expression by promoter replacement.