Difference between revisions of "Part:BBa J100303"
| (3 intermediate revisions by the same user not shown) | |||
| Line 4: | Line 4: | ||
The PmanP promoter, when induced by mannose, increases the beta-galactosidase activity of bacteria, which thus increases the expression of lacZ by four to seven-fold. In this experiment, we are using a 0.2% concentration of mannose in order to test the induction of transcription. | The PmanP promoter, when induced by mannose, increases the beta-galactosidase activity of bacteria, which thus increases the expression of lacZ by four to seven-fold. In this experiment, we are using a 0.2% concentration of mannose in order to test the induction of transcription. | ||
| + | |||
| + | |||
| + | ==Induction of pMAN promoter in <i>V. natriegens</i>== | ||
| + | ===(Characterized by iGEM Groningen-2019)<br>=== | ||
| + | |||
| + | The mannose inducible promoter is part of the pJOE8889 plasmid and controls the expression of the Cas9 enzyme. Since we designed all CRISPR components to work with this plasmid we also had to make sure that Cas9 would be expressed upon addition of mannose. There are no reports in the literature on the use of this promoter so we characterized it with the reporter mCherry. Moreover, <i>V. natriegens</i> cannot utilize mannose as a carbon source. Addition of 1 (w/v) % led to a 2.5 fold increase of expression within 12 hours. | ||
| + | |||
| + | [[File:BBa_J100303.png|500px|]] | ||
| + | |||
| + | Figure 1: pMAN is used in the pJOE8889 to control the expression of Cas9. In order to characterize this promoter we built it with mCherry and quantified the induction with mannose in <i>V. natriegens</i>. | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
Latest revision as of 22:07, 21 October 2019
PmanP
The PmanP promoter, when induced by mannose, increases the beta-galactosidase activity of bacteria, which thus increases the expression of lacZ by four to seven-fold. In this experiment, we are using a 0.2% concentration of mannose in order to test the induction of transcription.
Induction of pMAN promoter in V. natriegens
(Characterized by iGEM Groningen-2019)
The mannose inducible promoter is part of the pJOE8889 plasmid and controls the expression of the Cas9 enzyme. Since we designed all CRISPR components to work with this plasmid we also had to make sure that Cas9 would be expressed upon addition of mannose. There are no reports in the literature on the use of this promoter so we characterized it with the reporter mCherry. Moreover, V. natriegens cannot utilize mannose as a carbon source. Addition of 1 (w/v) % led to a 2.5 fold increase of expression within 12 hours.
Figure 1: pMAN is used in the pJOE8889 to control the expression of Cas9. In order to characterize this promoter we built it with mCherry and quantified the induction with mannose in V. natriegens.
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]