Assaying Promoter Activity Using LacZ and GFP as Reporters

  • Paul CarrollEmail author
  • Jade James
Part of the Methods in Molecular Biology book series (MIMB, volume 465)


The ability of bacteria to survive in a variety of different niches is due, in part, to their ability to respond and adapt to the environment. Extracellular signals are recognized by bacilli, and their responses are generally conducted at the transcript level. RNA polymerases recognize specific promoter regions on the genome and initiate transcription. Therefore, the analysis of gene expression is paramount to understanding the biology of an organism. In the case of pathogens, gene expression can alter during the course of the infection, and, therefore, specific targets can be identified for drug development. Promoter activity can be determined by cloning a promoter sequence upstream of a reporter gene and assaying the reporter activity, either from whole cells or from cell lysates. This chapter describes two reporter systems (GFP and LacZ) used for determining promoter activity that have been widely used in mycobacteria.


β-galactosidase cell-free extract fluorescence GFP LacZ live bacilli mycobacteria reporter vector 


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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  1. 1.Institute of Cell and Molecular Science, Barts and the London, Queen Mary’s School of Medicine and DentistryLondonUK
  2. 2.Centre for Infectious Disease, Institute of Cell and Molecular ScienceBarts and the London, Queen Mary’s School of Medicine and DentistryLondonUK

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