Difference between revisions of "Part:BBa J100303"

Revision as of 19:14, 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 Vibrio 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: Mannose inducible promoter in V. natriegens. Induction with 1% (w/v) mannose resulted in high expression of mCherry at 12 hours.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 9: / Line 9: @@
 ===(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 our 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.
+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.
 [[File:BBa_J100303.png|500px|]]
+Figure 1: Mannose inducible promoter in V. natriegens. Induction with 1% (w/v) mannose resulted in high expression of mCherry at 12 hours.