modified Lutz-Bujard LacO promoter,with alternative sigma factor σ32
Lac O and sigma 32 binding sites conserved sequences
modified Lutz-Bujard LacO promoter,with alternative sigma factor σ38
Lac O and Sigma 38 binding sites conserved sequences
modified Lutz-Bujard LacO promoter,with alternative sigma factor σ24
Lac O and sigma 24 binding conserved sequences
LacO and Sigma 24 binding conserved sequences
Lac O and sigma 38 binding sites conserved sequences
Lac O and sigma 38 binding sites conserved sequences.
modified Lutz-Bujard LacO promoter,with alternative sigma factor σ28
Lac O and sigma 28 binding sites' conserved sequences
Lac O and Sigma 28 binding sites' conserved sequences
Lac and sigma 28 conserved binding sites
Lac O and sigma 32 binding sites' promoter sequences
The universal stress protein A is a response of Escherichia coli cells to growth arrest, and its lacking generates cells with defective growth.This genetic element is dependent to sigma-70 factor. Its regulation is done through the concentration of a specific stationary phase allormone, guanosine-5'-diphosphate-3'-diphosphate (ppGp), as described elsewhere. The ppGp activate the transcription of downstream elements through a positive regulation of the β-subunit of RNA polymerase. In this sense, the PuspA element is considered a stationary phase promoter. However, its behavior is like a constitutive transcription promotion, becoming higher in stress situations. Diverse conditions make the Escherichia coli cell enter to stress state, like heat shocks, starvation, osmotic stress, ultraviolet light and other conditions. In these situations, the RNA polimerase sigma factors (ropS) trigger the expression of chaperones and other stress protector molecules, in order to help the cell survive. In our project we have used this promoter to trigger the expression of limonene synthase after an osmotic shock. Other uses of this part includes the environmental stress detectors making using a reporter system coupled to this promoter.
The original promoter site was shortened and the last three nucleotides were changed to restriction site adjustment.
We have drawn the sequence for an improved PuspA promoter from an analysis of 400bp upstream the Universal Protein A (UspA) gene in E. coli K12 MG16555 genome. After design, the DNA template of part was synthesized by IDT and was amplified from a G-block and assembled using 3A assembly method.