【摘要】 目的单克隆培养人神经母细胞瘤SH-SY5Y-细胞株,并鉴定其在基因水平缺失DHRS4基因簇,包括DHRS4、DHRS4L2和DHRS4L1的基因序列。方法有限稀释法培养实验室偶得的低表达DHRS4基因的人神经母细胞瘤SH-SY5Y细胞,相差显微镜观察细胞的生长和增殖,PCR、RT-PCR检测DHRS4、DHRS4L2和DHRS4L1基因的DNA和RNA含量,Westernblotting检测DHRS4蛋白的表达。结果筛选得到单克隆神经母细胞瘤细胞SH-SY5Y,在DNA、RNA和蛋白水平均未检测到DHRS4基因簇的DNA序列及表达产物。结论经过克隆低表达DHRS4基因簇的SH-SY5Y细胞,得到纯系的DHRS4基因簇缺失的SH-SY5Y-细胞株,该细胞对于进一步研究DHRS4基因簇的功能和表达调控都是难得的细胞模型。更多还原

【Abstract】 Objective To clone and identify the human neuroblastoma cell strain SH-SY5Y-with deletion of DHRS4 gene cluster,including the DHRS4,DHRS4L2 and DHRS4L1.Methods The human neuroblastoma cell line SH-SY5Y expressing low level of DHRS4 gene cluster were subcloned by limited dilution.The growth and proliferation of the cloned cells were observed by phase contrast microscope.PCR,RT-PCR and western-blotting were adopted to exam the DNA,RNA of DHRS4 gene cluster and protein of DHRS4.Results SH-SY5Y-cell strain were obtained by subcloning the human neuroblastoma cell line SH-SY5Y.There were no DNA,RNA and protein of DHRS4 gene cluster in SH-SY5Y-.Conclusion The cloned SH-SY5Y-cell strain are with deletion of DHRS4 gene cluster,thus provides a cell model for further studying the function and transcriptional regulation of DHRS4 gene cluster.更多还原