Study of Direct Interaction Between MiR30b and mRNA of Mouse Bach2
Duan Zhiqing
Li Yan
Wei Dinglu
Zhang Zhiwei
Abstract：Objective To analyze the novel target of miR30b in the B cell terminal differentiation.Methods Three target gene fragments,at lengths of 530,361 and 189 bp,were amplified from mouse cDNA by PCR using specific primers and site-direct mutant primers,respectively,and recovered with gel.The complete mutant fragment was obtained through jointing the 361 and 189 bp.The wild/mutant fragment of mouse Bach2 was inserted into pmirGLO vector,respectively to construct wild/mutant recombinanr luciferase reporter gene plasmid,pmirGLO-mBach2 and pmirGLO-mBach2 mt.They and miR30b co-transfected into HEK293 T cells.The Dual-Luciferase Reporter Assay System was performed to determine the Firefly-Renilla luciferase activity among groups.Results Restriction enzyme analysis and sequencing proved that recombinant plasmids,pmirGLO-mBach2 and pmirGLO-mBach2 mt,were constructed successfully.Compared with pmirGLO + miR30b group,luciferase activity in pmirGLO-mBach2 + miR30b group was significant decreased (P ＜ 0.05),while no significant change in pmirGLO-mBach2 mt + miR30b group (P ＞ 0.05).Conclusion Mouse Bach2 mRNA is a novel target gene of miR30b.
Keywords：miR30bmouse Bach2target geneB cell terminal differentiation
Publication Date：2017-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 84-87 )