Effect of HS2 and HS3 elements on erythroid-specific expression in transgenic mice  

作　　者：JIA Chunping, YAN Jingbin, XIAO Yanping, FANG Yudan, HUANG Shuzheng and ZENG Yitao(Institute of Medical Genetics, Shanghai Jiaotong University, Shanghai 200040, China) 

出　　处：《Progress in Natural Science:Materials International》2003年第9期672-677,共6页自然科学进展·国际材料（英文版）

基　　金：Supported by National 863 Hi-Tech Program of China (Grant No.2002AA206211) and the National Natural Science Foundation of China (Grant No.39893320)

摘　　要：The expression plasmids CMV/GFP, HS2ALL, HS3ALL and HS23ALL were selected to investigate the effect of HS2 and HS3 element on erythroid-specific expression in transgenic mice. These plasmids were digested with restriction enzymes and purified. And five DNA fragments, CMV/GFP, HS2/GFP, CMV/HS2/GFP, HS23/GFP and HS3/GFP were obtained. After purification, the above DNA fragments were microinjected into the pre-nuclei of the mice fertilized eggs and transgenic mice were generated, with an integration rate of 10.89%. The green fluorescence protein(GFP) expression in many transgenic mouse tissues was determined by FACS analysis. The results showed that the HS2 and 1.7 kb of β-globin gene promoter were sufficient for the erythroid-specific expression of β-globin gene. The GFP expression of different recombinant constructs was also analyzed in blood of all the transgenic mice with FACS. The results indicated that HS2 and HS3 had the same enhancement activity on the regulation of β-globin gene expression. Moreover, these two elements showed a significant synergistic effect on gene expression at the transgenic mouse level, although the GFP expression varied largely among different transgenic mouse litters.The expression plasmids CMV/GFP, HS2ALL, HS3ALL and HS23ALL were selected to investigate the effect of HS2 and HS3 element on erythroid-specific expression in transgenic mice. These plasmids were digested with restriction enzymes and purified. And five DNA fragments, CMV/GFP, HS2/GFP, CMV/HS2/GFP, HS23/GFP and HS3/GFP were obtained. After purification, the above DNA fragments were microinjected into the pre-nuclei of the mice fertilized eggs and transgenic mice were generated, with an integration rate of 10, 89% . The green fluorescence protein (GFP) expression in many transgenic mouse tissues was determined by FACS analysis. The results showed that the HS2 and 1. 7 kb of β-globin gene promoter were sufficient for the erythroid-specific expression of β-globin gene. The GFP expression of different recombinant constructs was also analyzed in blood of all the transgenic mice with FACS. The results indicated that HS2 and HS3 had the same enhancement activity on the regulation of β-globin gene expression. Moreover, these two elements showed a significant synergistic effect on gene expression at the transgenic mouse level, although the GFP expression varied largely among different transgenic mouse litters.

关 键 词：β-globin gene locus control region GFP transgenic mouse. 

分 类 号：Q789[生物学—分子生物学]