Fluorescent Reporters for investigating the Bacterial Stringent Response

摘要: The ``stringent response'' is a bacterial physiological adaptation to nutritional stress most well studied in the model organism \textit{Escherichia coli}. It research topic of interest both for increasing our understanding physiology general, and because potential informing development treatments diseases. small regulatory molecules penta- tetra-guanosine phosphate, collectively referred as (p)ppGpp, are known be central regulation stringent response, but convenient methods assessing intra-cellular concentrations (p)ppGpp currently lacking. One possible way assess intracellular levels molecule place ``reporter gene'' encoding fluorescent protein, under transcriptional control promoter regulated by molecule. Here, several plasmids variant green protein (GFP) different sequences this purpose have been constructed. promoters differ with respect whether how they effected level, previous evidence showing either postive (up-regulation), negative (down-regulation) or indifferent (no \textit{direct} regulation) response levels. Preliminary characterisation GFP production directed promoters, measured fluorescence, carried out cells growing non-perturbed conditions face elevated To elevate levels, fragment ppGpp synthase RelSeq from \textit{Streptococcus equisimilis} was induced use second plasmid.