Analysis of stochasticity in promoter activation by using a dual-fluorescence reporter system.
Stochastic dynamics of promoter activity in bacterial cells were studied by using a dual-fluorescence reporter system of protein expression. The dual-fluorescence reporter system enabled us to calculate the amplitude of intrinsic noise generated during transcription and translation. By fitting the experimental data to a simple stochastic model of protein expression, we could estimate parameters representing the stochastic transition between the active and inactive states of a promoter. Using the system, we analyzed the stochastic dynamics of promoter activation of genes in the lysine biosynthesis pathway in Escherichia coli. We found that the promoter of lysA has a significantly slower transition rate between active and inactive states than other promoters in the lysine biosynthesis pathway. The infrequent switching between active and inactive states can be a dominant source of noise in lysA expression. Analysis using the dual-fluorescence reporter system provided a better understanding of stochastic dynamics in promoter activation.
Ou
Jianhong
J
Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, Osaka, 565-0871, Japan.
Furusawa
Chikara
C
Yomo
Tetsuya
T
Shimizu
Hiroshi
H
D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics*; D002262:Carboxy-Lyases / Q000096:biosynthesis / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology*; D005434:Flow Cytometry; D005453:Fluorescence; D005786:Gene Expression Regulation / Q000502:physiology*; D008957:Models, Genetic; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology*; D013269:Stochastic Processes
D001426:Bacterial Proteins; D017931:DNA Primers; D002262:Carboxy-Lyases; C079912:LysA protein, Bacteria