Difference between revisions of "Part:BBa K5291040"
Kortybones (Talk | contribs) (→Usage and Biology) |
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===Usage and Biology=== | ===Usage and Biology=== | ||
− | Given the lack of RBS on pAB after zymotic digestion, we introduced pS, a universal promoter, into the plasmid. We synthesized | + | Given the lack of RBS on pAB after zymotic digestion, we introduced pS, a universal promoter, into the plasmid, as well as the other two promoters Pbla and Poprl. We synthesized them through the method of dimerization of positive and negative primers, and linked it to the plasmid backbone pAB. Then we screened for the three promoters by examining the fluorescence intensity seperately. We found that pS has the best effect of promotion, so we applied it to our plasmid.<br> |
<html><img width = "400" src="https://static.igem.wiki/teams/5291/images/part-wyn/ps-age-4.png" /></html> | <html><img width = "400" src="https://static.igem.wiki/teams/5291/images/part-wyn/ps-age-4.png" /></html> | ||
− | <br><b>Fig.1 The AGE figure of pS.</b><br> | + | <br><b>Fig.1 The AGE figure of pS.</b><br><br> |
+ | Then we cut off the GFP gene on pAB-pS by restriction endonuclease, gaining the usable backbone for reserved.<br><br> | ||
<html><img width = "400" src="https://static.igem.wiki/teams/5291/images/part-wyn/pab-ps.png" /></html> | <html><img width = "400" src="https://static.igem.wiki/teams/5291/images/part-wyn/pab-ps.png" /></html> | ||
<br><b>Fig.2 The AGE figure of pAB1-pS after digestion.</b><br><br> | <br><b>Fig.2 The AGE figure of pAB1-pS after digestion.</b><br><br> |
Revision as of 12:18, 1 October 2024
pS
The promoter pS is a kind of σ70-dependent constitutive promoter. It has been confirmed that pS could drive gene expression in wide range of host such as Pseudomonas putida and Azotobacter vinelandii.
Usage and Biology
Given the lack of RBS on pAB after zymotic digestion, we introduced pS, a universal promoter, into the plasmid, as well as the other two promoters Pbla and Poprl. We synthesized them through the method of dimerization of positive and negative primers, and linked it to the plasmid backbone pAB. Then we screened for the three promoters by examining the fluorescence intensity seperately. We found that pS has the best effect of promotion, so we applied it to our plasmid.
Fig.1 The AGE figure of pS.
Then we cut off the GFP gene on pAB-pS by restriction endonuclease, gaining the usable backbone for reserved.
Fig.2 The AGE figure of pAB1-pS after digestion.
By these ways, we can link the targeted genes to the vector and construct plasmids with functions.
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]