Abstract:Objective: To construct a luciferase reporter vector containing the 3′untranslated region (3′UTR) of Bcl-6 gene and measure the correlation between Bcl-6 and miR-346. Methods: The miRNA targeting Bcl-6 3′UTR was predicted by miRanda and PicTar. The synthetic 3′UTR fragment of Bcl-6 was cloned into PsiCHECK-2 reporter vector. The luciferase reporter vector and miRNA mimics were transfected into 293T cells. The relative luciferase activity was detected. Results: PicTar and miRanda database shared the results that miR346 have the complementary binding sites with 3′UTR of Bcl-6. Results of double enzyme digestion and DNA sequencing showed that sequence of luciferase reporter vector was correct.The luciferase activity of reporter vector treating miR346 was decreased observably about 66%. Conclusion: The luciferase reporter vector containing the 3′UTR of Bcl-6 was constructed successfully.
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