Cloning-free regulated monitoring of reporter and gene expression

BMC Mol Biol. 2009 Mar 8;10:20. doi: 10.1186/1471-2199-10-20.

Abstract

Background: The majority of the promoters, their regulatory elements, and their variations in the human genome remain unknown. Reporter gene technology for transcriptional activity is a widely used tool for the study of promoter structure, gene regulation, and signaling pathways. Construction of transcriptional reporter vectors, including use of cis-acting sequences, requires cloning and time-demanding manipulations, particularly with introduced mutations.

Results: In this report, we describe a cloning-free strategy to generate transcriptionally-controllable linear reporter constructs. This approach was applied in common transcriptional models of inflammatory response and the interferon system. In addition, it was used to delineate minimal transcriptional activity of selected ribosomal protein promoters. The approach was tested for conversion of genes into TetO-inducible/repressible expression cassettes.

Conclusion: The simple introduction and tuning of any transcriptional control in the linear DNA product renders promoter activation and regulated gene studies simple and versatile.

MeSH terms

  • Bacterial Proteins / genetics*
  • Carrier Proteins / genetics*
  • Cell Line
  • Cloning, Molecular
  • Gene Expression Regulation*
  • Genes, Reporter / genetics*
  • Humans
  • Models, Genetic

Substances

  • Bacterial Proteins
  • Carrier Proteins
  • Tet O resistance protein, Bacteria