Islet-1基因变异体的发现及其在神经干细胞内的表达

Discovery of rat Islet-1 gene variant and its expression in NSC

目的:构建大鼠Islet-1基因逆转录病毒表达载体,利用此载体将Islet-1基因转导入神经干细胞(NSC)内。方法:利用RT-PCR技术钓取大鼠Islet-1基因,将其插入到逆转录病毒载体plEGFP-C1中,利用包装细胞PA317将Islet-1基因转导入NSC内,观察Islet-1基因在NSC内的表达。结果:经PCR、酶切及荧光检测等证实,成功构建了plEGFP-C1-Islet-1表达载体,并应用免疫组化技术证明Islet-1在NSC内有表达。在实验中首次发现了Islet-1基因的变异体。结论:重组Islet-1基因逆转录病毒载体的构建为进一步探讨Islet-1基因是否参与NSC向运动神经元分化及其可能的作用机制奠定了坚实的实验基础。

AIM： To construct rat Islet-1 gene recombinant retroviral expression vector and to transduce Islet-1 gene into neural stem cell （NSC） by the vector. METHODS： The cDNA encoding the rat Islet-1 gene was isolated by RT-PCR method, the amplified gene fragment was subcloned into the retroviral vector plEGFP-C1. Islet-1 gene was transduced into NSC by PA317 packaging cells, then the expression of Islet-1 in NSC was observed. RESULTS： The recombinant Islet-1 retroviral vector was constructed successfully, as shown by PCR, restriction enzymes digestion and fluorescence detection, and the positive expression of Islet-1 was found in NSC by immunocytochemical staining. The variant of Islet-1 gene was also discovered in this study. CONCLUSION： The rat Islet-1 recombinant retroviral vector has been successfully prepared, which can be used to study the function of Islet-1 gene in NSC differentiation into cholinergic neurons.