Negative regulation role of SLC22A3 gene intron7 in gene transcription

Abstract:

Objective：To explore the regulatory effect of the human SLC22A3 gene intron7 and SNPrs2048327（A/G） on gene tran－scription. Methods：The hSLC22A3 intron7，amplified from the bacterial artificial chromosome（BAC） including whole human SLC22A3 gene，was cloned into pGL3-Basic and pGL3-Promoter respectively to construct the recombinant plasmid pGL3-Basic-SLCi7wt，pGL3-Promoter-SLCi7wt and pGL3-Promoter-SLCi7mut. The recombinants with internal reference plasmid pRL-SV40 were co-trans－fected into HEK293T cells，then the luciferase activity was detected. Results：There was no significant difference in luciferase activity between pGL3-Basic-SLCi7wt and pGL3-Basic（P=0.986）. Luciferase activity of pGL3-Basic-SLCi7wt was lower than that of pGL3-Promoter（（3.514 ± 0.096） vs. （6.286 ± 0.370）；P=0.000）. Luciferase activity of pGL3-Promoter-SLCi7mut was lower than that of pGL3-Promoter and pGL3-Promoter-SLCi7wt（P=0.000）. Conclusions：The human SLC22A3 gene intron7 has negative regulatory activity，which could significantly down-regulate the expression of luciferase reporter gene. The mutation（A→G） of SNPrs2048327（A/G） located in the hSLC22A3 intron7 may enhance the negative regulatory activity of this intron.