Difference between revisions of "Part:BBa K5291040"
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Given the lack of RBS sequence on pAB1 after zymotic digestion, we introduced pS, a universal promoter, into the plasmid. We synthesized it through the method of dimerization of positive and negative primers, and linked it to the plasmid backbone pAB1.<br> | Given the lack of RBS sequence on pAB1 after zymotic digestion, we introduced pS, a universal promoter, into the plasmid. We synthesized it through the method of dimerization of positive and negative primers, and linked it to the plasmid backbone pAB1.<br> | ||
| − | <html><img width = " | + | <html><img width = "250" 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> | <br><b>Fig.1 The AGE figure of pS.</b><br><br> | ||
| − | Then we cut off the GFP gene on pAB1-pS by restriction endonuclease, gaining the usable backbone for reserved. | + | Then we cut off the GFP gene on pAB1-pS by restriction endonuclease, gaining the usable backbone for reserved.<br> |
| − | <html><img width = " | + | <html><img width = "250" 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> | ||
| − | By these ways, we can link the targeted genes to the vector and | + | By these ways, we can link the targeted genes to the vector.<br><br> |
| − | + | Later, we conducted the effect verification of pS. We put the plates with <i>Pseudomonas aeruginosa</i> under UV and intense fluorescence has been observed, which means that pS is able to come into effect in PAO1. <br><br> | |
| + | <html><img width = "500" src="https://static.igem.wiki/teams/5291/images/part-wyn/ps-effect-fluorescence.jpg" /></html> | ||
| + | <br><b>Fig.3 The plates with glowing PAO1 under UV.</b><br><br> | ||
| + | In conclusion, we successfully constructed the plasmid with the effective promoter. | ||
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Latest revision as of 12:14, 2 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 sequence on pAB1 after zymotic digestion, we introduced pS, a universal promoter, into the plasmid. We synthesized it through the method of dimerization of positive and negative primers, and linked it to the plasmid backbone pAB1.
Fig.1 The AGE figure of pS.
Then we cut off the GFP gene on pAB1-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.
Later, we conducted the effect verification of pS. We put the plates with Pseudomonas aeruginosa under UV and intense fluorescence has been observed, which means that pS is able to come into effect in PAO1.
Fig.3 The plates with glowing PAO1 under UV.
In conclusion, we successfully constructed the plasmid with the effective promoter.
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]