摘要
目的构建人工miRNA表达簇,并研究miRNA的加工对蛋白质表达水平的影响。方法从miRNA表达库中扩增miR-34a及miR-424的前体序列,将上述两种miRNA串联,构建PGL-34a-424人工miRNA簇表达载体。通过RT-PCR的方法检测人工miRNA簇的表达情况。然后将miRNA前体突变,再用荧光素酶报告基因技术研究miRNA的加工对蛋白质表达的影响。结果成功构建人工miRNA簇表达载体,荧光素酶实验显示miRNA的加工能够抑制蛋白的表达。结论人工miRNA表达簇能够高效表达,miRNA的加工对蛋白质的表达有抑制作用。
Objective To construct artificial microRNA cluster expression vector and a preliminary study on the relation between miRNA procession and protein expression.Methods The recombinant plasmids （PGL-34a-424 ） were constructed by amplifing miR-34a and miR-424 precursors from a library of vectors expressing miRNAs and connecting them.The expression of artificial microRNA cluster was tested by RT-PCR.Then,the relation between miRNA procession and protein expression was detected by Luciferase Reporter Assay after constructing mutant miRNA precursors.Results Artificial microRNA cluster expression vector was constructed successfully and miRNA procession could suppress protein expression by Luciferase Reporter Assay.Conclusion Artificial microRNA cluster can efficiently express,and miRNA procession can suppress protein expression.
出处
《安徽医科大学学报》
CAS
北大核心
2010年第3期294-297,共4页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金资助项目(编号:30873008)
