Difference between revisions of "Part:BBa K2972013"
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| + | <h1>Characterization of Promoter</h1> | ||
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| + | Construction of gene expressing vector: We connected PT7 and mcherry to the plasmid pACYCDute-1. Then we characterized promoter intensity through fluorescence intensity.<br><br> | ||
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| + | Experiment:<br> | ||
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| + | The plasmid was transformed to E.coli BL21, and then we transferred the E. coli introduced into the plasmid into LB medium containing chloramphenicol successfully. After 12 hours of culture, it was transferred to 40 ml LB medium containing chloramphenicol with initial OD600 of 0.01. After 5 hours of transfer of E.coli containing T7 promoter, 1/1000 IPTG was added, and samples were taken every 2 hours for a total of 12 hours.<br><br> | ||
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| + | The results are shown below:<br> | ||
[[File:BIT-China silver 2.png|center|thumb|400px|<b>Fig.Fluorescence fermentation results of T7 promoter.</b>]] | [[File:BIT-China silver 2.png|center|thumb|400px|<b>Fig.Fluorescence fermentation results of T7 promoter.</b>]] | ||
Latest revision as of 02:30, 22 October 2019
T7 Promoter Test Composition
Characterization of Promoter
Construction of gene expressing vector: We connected PT7 and mcherry to the plasmid pACYCDute-1. Then we characterized promoter intensity through fluorescence intensity.
Experiment:
The plasmid was transformed to E.coli BL21, and then we transferred the E. coli introduced into the plasmid into LB medium containing chloramphenicol successfully. After 12 hours of culture, it was transferred to 40 ml LB medium containing chloramphenicol with initial OD600 of 0.01. After 5 hours of transfer of E.coli containing T7 promoter, 1/1000 IPTG was added, and samples were taken every 2 hours for a total of 12 hours.
The results are shown below:
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 611
Illegal AgeI site found at 723 - 1000COMPATIBLE WITH RFC[1000]