Construction and functional analysis of luciferase reporter plasmid containing connexin43 gene promoter

Abstract

Connexin 43 (Cx43), the principal gap junction protein in vascular smooth muscle cells(VSMCs) resulting in modulation of gap junction. Myocardin is thought to have a key role in Vascular Smooth Muscle Cell (VSMC) development by acting on CArG-dependent genes. In this study, a human Cx43 promoter luciferase reporter construct was generated by PCR amplification of Cx43 promoter. The PCR fragment was digested and cloned into pGL3-Basic vector. The promoter sequence was verified by sequencing. The results showed that luciferase reporter plasmids of Cx43 promoter were successfully constructed. Then the effects of a key transcription factor, which plays important roles in Cx43 levels control, were investigated by luciferase reporter assays in HEK293. The results showed that myocardin can enhance transcriptional activity of Cx43. Our observations provide evidence that Myocardin targets Cx43 expression directly

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