Construction and Functional Analysis of Luciferase Reporter Plasmid Containing GSNOR Gene Promoter
D.L. Zheng, X.H. Liao, Z.Q. Wei, F. Du, H.P. He, T.C. Zhang
Available Online March 2016.
- https://doi.org/10.2991/meep-15.2016.51How to use a DOI?
- GSNO; GSNOR; S-nitrosylation; luciferase activity assay
- 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.
- Open Access
- This is an open access article distributed under the CC BY-NC license.
Cite this article
TY - CONF AU - D.L. Zheng AU - X.H. Liao AU - Z.Q. Wei AU - F. Du AU - H.P. He AU - T.C. Zhang PY - 2016/03 DA - 2016/03 TI - Construction and Functional Analysis of Luciferase Reporter Plasmid Containing GSNOR Gene Promoter BT - International Conference on Materials Chemistry and Environmental Protection 2015 PB - Atlantis Press SP - 194 EP - 197 SN - 2352-541X UR - https://doi.org/10.2991/meep-15.2016.51 DO - https://doi.org/10.2991/meep-15.2016.51 ID - Zheng2016/03 ER -