βLCR对β地中海贫血基因在转基因小鼠体内表达的影响

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2类转基因鼠中外源基因的表达量。结果：成功建立2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核心片段的存在可以使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。

Effects of βLCR on β-thalassaemia gene expression in transgenic mice

The thalassemia is a heterogeneous group of inherited disorders. The main pathological mechanism which causes thalassemia is the imbalance in the synthesis of αandβglobin chains, due to mutations in the globin loci. The C→T transition at nucleotide 654 of intron 2 inβglobin gene is one of the most frequent βthalassemia mutation(β-IVSⅡ-654) 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：We wanted to identify that what contribution wouldβLCR give toβ thalassemia gene (β-IVSⅡ-654) in the transgenic mouse. Methods: To prepare two genes, one was the merelyβthalassemia gene, the other was a modified βthalassemia 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 merelyβthalassemia gene was more lower than the modified βthalassemia gene contains βLCR in the transgenic mouse. Conclusion: We therefore conclude that a combination ofβLCR is necessary to highly expressionsβthalassemia gene in the transgenic mouse.