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title: Construction and Functional Analysis of Luciferase Reporter Plasmid Containing GSNOR Gene Promoter publication: meep-15 part of series: Advances in Physics Research ISBN: 978-94-6252-169-8 ISSN: 2352-541X DOI: doi:10.2991/meep-15.2016.51 (how to use a DOI) author(s): D.L. Zheng, X.H. Liao, Z.Q. Wei, F. Du, H.P. He, T.C. Zhang corresponding author: D.L. Zheng publication date: March 2016 keywords: GSNO; GSNOR; S-nitrosylation; luciferase activity assay abstract: S-nitrosoglutathione (GSNO) is a nitrosylating agent which mediates the posttranslational process of Snitrosylation on proteins resulting in modulation of their activity. GSNOR (S-Nitrosoglutathione reductase) is an enzyme related to GSNO degration, which has been used extensively in controlling S-nitrosylation levels in vivo. 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 GSNOR promoter luciferase reporter construct was generated by PCR amplification of GSNOR promoter. The PCR fragment was digested and cloned into pGL3 vector. The promoter sequence was verified by sequencing. The results showed that luciferase reporter plasmids of GSNOR promoter were successfully constructed. Then the effects of a key transcription factor, which plays important roles in S-nitrosylation levels control, were investigated by luciferase reporter assays in VSMCs. The results showed that myocardin can enhance transcriptional activity of GSNOR. Our research will help us to screen some novel factors in regulating S-nitrosylation levels in vivo. @Monash2011 |
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