Assaying promoter activity using LacZ and GFP as reporters.
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.
Carroll
Paul
P
Institute of Cell and Molecular Science, Barts and the London, Queen Mary's School of Medicine and Dentistry, 4 Newark Street, Whitechapel, London E1 2AT, UK. p.carroll@qmul.ac.uk
James
Jade
J
D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics*; D007763:Lac Operon; D009161:Mycobacterium / Q000235:genetics*; D011401:Promoter Regions, Genetic; D001616:beta-Galactosidase / Q000235:genetics
D049452:Green Fluorescent Proteins; D001616:beta-Galactosidase