Promega Corporation

Improve Promoter Analysis with Luciferase Reporters

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The pGL4.10 [luc2] Vector (Cat. #E6651) contains a multiple cloning site for insertion of promoter sequences.

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The pGL4.74 [hRluc/TK] Vector (Cat. #E6921) ready to use as a control vector in Dual-Luciferase® Reporter experiments.

  • Promoter analysis seeks to understand why a particular gene responds to a particular environmental condition or treatment.
  • Originally, promoter dissection involved deletion analysis of the proximal promoter region until the gene response to the condition or treatment changed or was eliminated.
  • Today, bioinformatics allows researchers to make "educated guesses" about what regions of the promoter will be important for a particular response.
  • Promoter analysis is being used to ask broad questions, like why do certain polymorphisms persist in the genome, what makes a primate different from a human when the genes are identical, etc.
  • The pGL4 vectors represent the latest technology for promoter analysis.
  • We dramatically reduced background by removing hundreds of potential transcription factor binding sites from the vector backbone.
  • The re-engineered luciferase gene offers unparalleled sensitivity because the signal produced from the luciferase reaction is 200-fold brighter signal than that from the native luciferase gene.

Long Terminal Repeat Promoter Analysis of HIV-1

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Treating T-cells infected with HIV-1 involves activating the provirus that has integrated into the genome so that these cells can be "marked" for treatment. However, activating the provirus triggers the NF-κB pathway and activates the T-cell, allowing the virus to replicate and infect additional cells. The authors of this study screened for factors that could activate a HIV-1 promoter in which the NF-κB sites were mutated.

Yang, H-C. et al. (2009) PNAS 106, 6321–6.

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