Difference between revisions of "Part:BBa K4193026"
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− | + | Cytokinin isopentenyladenine (IP), as quorum sensing molecule, is the product of ATP catalyzed by AtlPT4. When the signal molecule IP reaches a certain concentration, it activates the SSRE promoter and increases the expression of production gene controlled by SSRE promoter in our experiment, to better characterize SSRE promoter and quorum sensing circuit, we put eGFP gene at the downstream of SSRE promoter. | |
+ | <html> | ||
+ | <head> | ||
+ | <meta charset="utf-8"> | ||
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+ | </head> | ||
+ | <body> | ||
+ | <div> | ||
+ | <img src="https://static.igem.org/mediawiki/parts/1/17/QS.jpg" style="width: 80%;"> | ||
+ | </div> | ||
+ | </body> | ||
+ | </html> | ||
+ | Figure1.The signal pathway of IP-mediated quorum sensing | ||
+ | We successfully introduced cytokinin-mediated quorum sensing circuit into the genome of Aureobasidium melanogenum P16. To characterize this circuit and SSRE promoter, we added different concentration of IP and introduced eGFP at the downstream of SSRE promoter. We measured the fluorescence to figure out the dynamic regulation range. | ||
+ | <html> | ||
+ | <head> | ||
+ | <meta charset="utf-8"> | ||
+ | |||
+ | </head> | ||
+ | <body> | ||
+ | <div> | ||
+ | <img src="https://static.igem.wiki/teams/4193/wiki/model/qs-result.jpg" style="width: 80%;"> | ||
+ | </div> | ||
+ | </body> | ||
+ | </html> | ||
+ | Figure2.Gel electrophoresis results of P16 genome amplifying AtCRE1 gene (Lane1), transformed strain genome amplifying AtCRE1 (Lane2), P16 genome amplifying PTP2+eGFP (Lane3) and transformed strain genome amplifying PTP2+eGFP (Lane 4) | ||
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+ | Different concentrations of IP can increase the expression of SSRE promoter. After rough statistical analysis, the expression intensity of SSRE promoter was significantly correlated with IP concentration. | ||
+ | <html> | ||
+ | <head> | ||
+ | <meta charset="utf-8"> | ||
+ | |||
+ | </head> | ||
+ | <body> | ||
+ | <div> | ||
+ | <img src="https://static.igem.wiki/teams/4193/wiki/other/part/ssre-expression.jpg"> | ||
+ | </div> | ||
+ | </body> | ||
+ | </html> | ||
+ | Figure3. Effect of different concentrations of IP on SSRE promoter induction. | ||
+ | |||
+ | To get a great insight for the protein in designed signal pathway, we used dry lab to make up for the defect we could not characterize the behavior of these proteins, such as AtCRE1_P, Ypd1_P and Skn7_P interacting with different subtypes of SSRE promoter. | ||
+ | <html> | ||
+ | <head> | ||
+ | <meta charset="utf-8"> | ||
+ | |||
+ | </head> | ||
+ | <body> | ||
+ | <div> | ||
+ | <img src="https://static.igem.wiki/teams/4193/wiki/other/part/phophoylated-function-protein-changes.jpg"> | ||
+ | </div> | ||
+ | </body> | ||
+ | </html> | ||
+ | Figure4.The behavior of proteins in designed signal pathway | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== |
Latest revision as of 12:08, 12 October 2022
QS system
Cytokinin isopentenyladenine (IP), as quorum sensing molecule, is the product of ATP catalyzed by AtlPT4. When the signal molecule IP reaches a certain concentration, it activates the SSRE promoter and increases the expression of production gene controlled by SSRE promoter in our experiment, to better characterize SSRE promoter and quorum sensing circuit, we put eGFP gene at the downstream of SSRE promoter.
We successfully introduced cytokinin-mediated quorum sensing circuit into the genome of Aureobasidium melanogenum P16. To characterize this circuit and SSRE promoter, we added different concentration of IP and introduced eGFP at the downstream of SSRE promoter. We measured the fluorescence to figure out the dynamic regulation range.
Different concentrations of IP can increase the expression of SSRE promoter. After rough statistical analysis, the expression intensity of SSRE promoter was significantly correlated with IP concentration.
To get a great insight for the protein in designed signal pathway, we used dry lab to make up for the defect we could not characterize the behavior of these proteins, such as AtCRE1_P, Ypd1_P and Skn7_P interacting with different subtypes of SSRE promoter.
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1092
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Illegal PstI site found at 6143 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1092
Illegal EcoRI site found at 3483
Illegal EcoRI site found at 5357
Illegal EcoRI site found at 5682
Illegal SpeI site found at 5194
Illegal PstI site found at 5444
Illegal PstI site found at 6143 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1092
Illegal EcoRI site found at 3483
Illegal EcoRI site found at 5357
Illegal EcoRI site found at 5682
Illegal BglII site found at 523
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Illegal BamHI site found at 633 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1092
Illegal EcoRI site found at 3483
Illegal EcoRI site found at 5357
Illegal EcoRI site found at 5682
Illegal XbaI site found at 2581
Illegal XbaI site found at 3040
Illegal XbaI site found at 3124
Illegal XbaI site found at 3487
Illegal XbaI site found at 6643
Illegal SpeI site found at 5194
Illegal PstI site found at 5444
Illegal PstI site found at 6143 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1092
Illegal EcoRI site found at 3483
Illegal EcoRI site found at 5357
Illegal EcoRI site found at 5682
Illegal XbaI site found at 2581
Illegal XbaI site found at 3040
Illegal XbaI site found at 3124
Illegal XbaI site found at 3487
Illegal XbaI site found at 6643
Illegal SpeI site found at 5194
Illegal PstI site found at 5444
Illegal PstI site found at 6143
Illegal NgoMIV site found at 6410
Illegal NgoMIV site found at 6450
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