Difference between revisions of "Part:BBa K4583000"

Revision as of 14:45, 4 October 2023

PYU3

PYU3 is the promoter of the gene orf-0464, and it comes from Escherichia coli. It will reach its maximum expression at the late stationary phase.

Usage and Biology

When the bacteria enter the stationary phase, the physiological state of the bacteria changes significantly. During this phase, many genes will respond to make timely adjustments. This part (BBa_K4583000) is the promoter of the gene orf-0464. Its most notable feature is that it will be expressed in the late stationary phase. Sequence and Features

Characterization

Protocols

Our experimental conditions for characterizing this part were as follows:

• E. coli MG1655
• 30^oC, 48h, under vigorous shaking
• Plasmid Backbone: PACYC
• Equipment: Multi-Detection Microplate Reader (Synergy HT, Biotek, U.S.)

We used GFP (excitation at 485 nm and emission at 528 nm)and BFP (excitation at 400 nm and emission at 450 nm) to characterize this part. As our focus was mainly on the expression time, we processed the obtained fluorescence data by means of the following equation: x'=(x-min)/(max-x). This treatment makes all data fall between 0 and 1, which is easier to use for comparisons between different fluorescence data (since our focus is on expression time).

Characterization using fluorescent proteins

[[Image:pyu3l19l31.png|800px|thumb|center|Figure 2: Fluorescence per cell of yeast cells transformed with a plasmid carrying BBa_K801020 in front of an eGFP-gene. The different alcohol concentrations are indicate

Comparation with another promoter

Comparison of expression time with cell density

Reference