摘要
目的:通过双荧光素酶报告系统验证miR-6857-3p对SP110基因的靶向调控作用。方法:从人巨噬细胞基因组中获取SP110基因的3′UTR序列,构建SP110基因的3′UTR野生型及SP110的3′UTR突变型双荧光素酶报告基因载体,分别将双荧光素酶报告质粒(psi CHECK-2-SP110-3′UTR和psi CHECK-2-SP110-3′UTR-6857mut)和miRNA质粒(miR-6857-3p mimic、miR-4772-3p mimic、miR-4659b-3p mimic和miRNA mimic nc)共转染到293T细胞,进行荧光活性测定。结果:与对照组相比,携带miR-6857-3p mimic和SP110野生型3'UTR的构建体的相对荧光素酶活性产生明显下降(P<0.001),SP110突变型3'UTR的构建体的相对荧光素酶活性下降没有统计学意义(P>0.05)。结论:miR-6857-3p可以直接靶向SP110的3′UTR序列,SP110是miR-6857-3p的直接靶基因。
Objective:To verify the targeting regulation of miR-6857-3 p on SP110 gene by dual luciferase reporter system.Methods:The 3’UTR sequence of SP110 was obtained from the gene bank,and the 3’UTR wild type of SP110 and the 3’UTR mutant double luciferase reporter gene vector of SP110 were constructed.The dual luciferase reporter plasmid(psi CHECK-2-SP110-3′UTR and psi CHECK-2-SP110-3′UTR-6857 mut)and the miRNA plasmid(miR-6857-3 p mimic,miR-4772-3 p mimic,miR-4659 b-3 p mimic and miRNA mimic nc)were co-transfected into 293 T cells for fluorescence activity assay.Results:Compared with the control group,the relative luciferase activity of the construct carrying miR-6857-3 p mimic and SP110 wild type 3’UTR was significantly decreased(P<0.001).The relative luciferase activity of the SP110 mutant 3’UTR construct was not statistically significant(P>0.05).Conclusion:miR-6857-3 p can directly target the 3’UTR sequence of SP110,SP110 is a direct target gene of miR-6857-3 p.
作者
李新月
王雪
贾鹏霞
张万江
程江
吴江东
马雅静
LI Xin-yue;WANG Xue;JIA Peng-xia;ZHANG Wan-jiang;CHENG Jiang;WU Jiang-dong;MA Ya-jing(Shihezi University School of Medicine,Xinjiang Shihezi,823002;Key Laboratory of Xinjiang Endemic and Ethnic Diseases,Shihezi University,Xinjiang Shihezi,832002;Department of Clinical Laboratory,the First Affiliated Hospital of Shihezi University School of Medicine,Xinjiang Shihezi,832008)
出处
《农垦医学》
2020年第2期97-100,共4页
Journal of Nongken Medicine
基金
NSFC-新疆联合基金资助项目(U1803127)
兵团重点领域科技攻关计划项目(2018AB019)。
