Difference between revisions of "Part:BBa K2972009"
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<h1>Characterization of Promoter</h1> | <h1>Characterization of Promoter</h1> | ||
| − | Construction of gene expressing vector: We connected PsigmaB and mcherry to the plasmid pACYCDute-1. Then we characterized promoter intensity through fluorescence intensity.<br> | + | Construction of gene expressing vector: We connected PsigmaB and mcherry to the plasmid pACYCDute-1. Then we characterized promoter intensity through fluorescence intensity.<br><br> |
Experiment:<br> | Experiment:<br> | ||
| − | 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. Samples of E.coli containing PsigmaB were taken every two hours. 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> | + | 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. Samples of E.coli containing PsigmaB were taken every two hours. 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> |
The results are shown below:<br> | The results are shown below:<br> | ||
| + | [[File:BIT-China-余凡尘-银奖part PσB.png|thumb|center|400px|<b>Fig.Fluorescent fermentation results of BBa_K2972009.</b>]]<br><br> | ||
Revision as of 20:47, 21 October 2019
SigmaB Promoter Test Composition
Sequence and Features
- 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 759
Illegal AgeI site found at 871 - 1000COMPATIBLE WITH RFC[1000]
Characterization of Promoter
Construction of gene expressing vector: We connected PsigmaB 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. Samples of E.coli containing PsigmaB were taken every two hours. 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:
