Abstract：Objective: To identify the targetedregulating relationship between miRNA-625 and AKT2 via constructing luciferase reporter gene vector. Methods: The AKT2 was predicted as miRNA-625 target gene by biological software. The 3′-untranslated regions(3′-UTR) of the wild type AKT2 and mutant AKT2 sequence were cloned into luciferase reporter vector pMIR-Report. The HEK-293T cells were divided into four groups randomly, wild-type plasmid + miRNA625 group, wild-type plasmid + miRNA negative control group, mutant plasmid + miRNA-625 group, mutant plasmid+miRNA negative control group.The relevant plasmids and miRNAs were transfected into groups by Lipofectamine 2000. The luciferase activity was detected by dual luciferase reporter gene system. Results: The recombinant plasmids were identified correctly. Dual-luciferase reporter assay system revealed luciferase activity of wildtype plasmid+miRNA-625 group was significantly lower than that of wild-type plasmid+ miRNA negative control group(P<0.05). Conclusion: The miRNA-625 inhibited the luciferase activity of AKT2 wild type vector, which indicates AKT2 gene could be targeted regulated by miRNA-625.
收稿日期: 2016-02-29
基金资助:
国家自然科学基金资助项目（81370119)；镇江市社会发展项目（SH2015044)
通讯作者:
钱粉红（通讯作者)，副主任医师，E-mail:zhaoqian604@126.com
作者简介: 魏斌（1990—)，男，硕士研究生
引用本文:
魏斌, 邓霞, 卞秀娟. AKT2 3′-UTR荧光素酶报告基因载体构建及与miRNA-625靶向关系验证[J]. 江苏大学学报:医学版, 2016, 26(03): 204-.
WEI Bin, Deng-Xia, Bian-Xiu-Juan. Construction of the AKT2 3′-UTR luciferase reporter gene vector and verification of the targeted relationship with miRNA-625. Journal of Jiangsu University(Medicine Edition), 2016, 26(03): 204-.
2016, Vol. 26 
