Biotechnology Letters

, Volume 34, Issue 5, pp 823–830 | Cite as

A dual color fluorescent reporter system for the real time detection of promoter activity

  • Liushuai Hua
  • Mingxun Li
  • Xiaomei Sun
  • Jing Wang
  • Zhuanjian Li
  • Yao Xu
  • Shenrong Hu
  • Hong Chen
Original Research Paper

Abstract

Understanding the mechanisms controlling transcription of a gene requires the identification and characterization of its cis-acting regulatory elements. A highly useful approach to the identification and characterization of cis-acting elements has been the systematic coupling of genomic fragments to reporter constructs, so called “promoter bashing”. The expression from such reporters must be normalized for differences in transient transfection efficiency between cells and replicates. A novel dual color fluorescent reporter system to assay the promoter activity of a genomic DNA fragment of interest was established by cloning a Discosoma red fluorescent protein gene and a green fluorescent protein gene into a single vector, giving a system in which the ratio between red and green fluorescence is proportional to promoter activity. This system allows real time quantitative monitoring of promoter activity. We validated this approach by assaying the cis-acting regulatory potential of the peroxisome proliferators-activated receptor gamma2 gene.

Keywords

Discosoma red protein Dual color fluorescent reporter system Green fluorescent protein Peroxisome proliferators-activated receptor gamma2 gene Promoter activity 

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Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Liushuai Hua
    • 1
  • Mingxun Li
    • 1
  • Xiaomei Sun
    • 1
  • Jing Wang
    • 1
  • Zhuanjian Li
    • 1
  • Yao Xu
    • 1
  • Shenrong Hu
    • 1
  • Hong Chen
    • 1
  1. 1.College of Animal Science and Technology, Shaanxi Key Laboratory of Molecular Biology for AgricultureNorthwest A&F UniversityYangling, ShaanxiChina

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