人β-珠蛋白MAR对转基因在不同细胞中表达的影响

Effect of human β-globin MAR on transgene expression in different cells

目的:研究人β珠蛋白核基质结合区(matrixattachmentregion,MAR)在不同表达宿主中对转基因表达的影响.方法:通过PCR从人基因组扩增β珠蛋白MAR,正向克隆至pCAT3control载体SV40启动子的上游,分别检测在瞬时及稳定表达的情况下,MAR在NIH3T3及Hela细胞中对CAT报告基因的影响情况.结果:在瞬时表达情况下,NIH3T3细胞及Hela细胞中MAR均不能提高CAT基因的表达;在稳定整合的情况下,NIH3T3细胞中β珠蛋白MAR可使CAT报告基因表达水平提高8倍,而在Hela细胞表达水平只提高2.5倍.结论:MAR能在一定程度上提高稳定外源基因的表达水平;MAR的作用与表达宿主相关.

AIM: To study the effect of human β-globin matrix attachment region (MAR) on transgene expression in different host cells. METHODS: The β-globin MAR was amplified by PCR and cloned into the upstream of SV40 promoter of pCAT-control vector in forward orientation and the CAT reporter gene expression was examined under transient and stable expression respectively. RESULTS: The results showed that there was little effect of MAR on CAT transgene expression in transient expression. Under stable expression, the β-globin MAR could enhance the CAT gene expression 8 folds in NIH3T3 cells, but only 2 folds in Hela cells. CONCLUSION: The effect of MAR on transgene expression is related to the host cells.