U2AF65蛋白表达水平影响基因UBQLN1的可变剪接  

Changes of U2AF65 Expression Level Affect Alternative Splicing of UBQLN1 Gene

作　　者：瞿秀华[1] 刘萱[1] 李晓明[1] 李平[1] 马清钧[1] 曹诚[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100850

出　　处：《生物技术通讯》2010年第4期470-473,共4页Letters in Biotechnology

摘　　要：目的:研究U2AF65蛋白的表达水平对基因UBQLN1可变剪接的影响。方法:应用pSR-GFP/Neo载体构建2个U2AF65-siRNA干扰载体,转染293T细胞,通过Western印迹、QRT-PCR检测干扰效果,RT-PCR验证基因UBQLN1的可变剪接。结果:利用设计的U2AF65-siRNA能够干扰细胞中U2AF65的表达;RT-PCR结果显示U2AF65表达水平的下降促使UBQLN1第8外显子的跳跃增加。结论:U2AF65可以通过表达水平的变化参与调控基因UBQLN1的可变剪接。Objective： To investigate the effect of U2AF65 expression level on alternative splicing of UBQLN1 gene. Methods： Two U2AF65-siRNA vectors were constructed by using the pSR-GFP / Neo vector, and then they were transfected into 293T cells. The effect of RNA interference was examined by methods of Western blotting and QRT-PCR, and the alternative splicing of UBQLN1 was detected through RT-PCR. Results： The expression of U2AF65 was suppressed by U2AF65-siRNA vectors. The result of RT-PCR revealed that knockdown of U2AF65 led to an increase in exon 8 skipping of UBQLN1. Conclusion： U2AF65 may regulate alternative splicing of UBQLN1 through changes of expression level.

关 键 词：U2AF65蛋白 RNA干扰 UBQLN1基因 可变剪接 

分 类 号：Q75[生物学—分子生物学]