βLCR对β地中海贫血基因在转基因小鼠体内表达的影响

Effects of βLCR on β-thalassaemia Gene Expression in Transgenic Mice

目的:明确在转基因小鼠体内,βLCR对β地中海贫血基因表达的影响。方法:将完整人β-IVSⅡ-654地中海贫血基因,与串连了人βLCR的β-IVSⅡ-654地中海贫血基因分别经显微注射法制作转基因小鼠;荧光定量RT-PCR法检测β-IVSⅡ-654地贫基因在转基因小鼠体内的表达;采用统计分析比较两类转基因鼠中外源基因的表达量。结果:成功建立两类整合了人β-IVSⅡ-654地贫基因的转基因小鼠模型。荧光定量RT-PCR分析结果表明,在整合了串连人βLCR的β-IVSⅡ-654地贫基因的小鼠体内,外源基因mRNA的表达量远高于仅整合β-IVSⅡ-654地贫基因的小鼠(统计分析P值)。结论:βLCR核心片段的存在是β珠蛋白基因家族(包括β-地贫基因)在转基因小鼠体内获得高效表达的必要条件。

The thalassemia is a heterogeneous group of inherited disorders. The main pathological mechanism which causes thalassemia is the imbalance in the synthesis of α and β globin chains, due to mutations in the globin loci. The C→T transition at nucleotide 654 of intron 2 in β globin gene is one of the most frequent β thalassemia mutation（ β-IVS Ⅱ 4554） in GuangDong China. The expression of human β globin gene cluster is definitely regulated with strict developmental stage and tissue lineage specificities as well as the delicated control of the biosynthetic equilibrium. It was all know that the expression of human β- globin gene cluster concerned with the β locals control region（βLCR） in human being. Purpose ： The contribution that βLCR give to β thalassemia gene （ β-IVS Ⅱ 4554） in the transgenic mouse was wanted to be identified. Methods： To prepare two genes, one was the merely β thalassemia gene, the other was a modified β thalassemia gene contains βLCR. To made up two lines of transgenic mouse by use these two genes. Then, detect the expression of two genes in the two transgenic mouse lines by fluorescent quantitation RT- PCR. To compared the two expressions of two genes by statistical analysis. Result： Two transgenic mouse were established. The statistical analysis shows that the expression of the merely β thalassemia gene was more lower than the modified β thalassemia gene contains βLCR in the transgenic mouse. Conclusion： A combination of βLCR is necessary to highly expressions of β thalassemia gene in the transgenic mouse.