Difference between revisions of "Part:BBa K4583000"
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===Characterization using fluorescent proteins=== | ===Characterization using fluorescent proteins=== | ||
| + | ===CComparation with another promoter=== | ||
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===Comparison of expression time with cell density=== | ===Comparison of expression time with cell density=== | ||
==Reference== | ==Reference== | ||
Revision as of 14:16, 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Characterization
Protocols
Our experimental conditions for characterizing this part were as follows:
- E. coli MG1655
- 30oC, 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).