Part:BBa_K5210018
PsacB-SOD-catalase
We established a sucrose-sensing switch in Vibrio natriegens. This composite part is regulated by the promoter PsacB , which can be activated in the presence of sucrose to express the downstream SOD, catalase to catabolise ROS.
Usage and Biology
Since the effect of pSacB promoter is more clear, our experiment mainly explored whether SOD and catalase can be expressed.Pet-28a was used as a vector for two target genes, and the proteins were expressed in engineering bacteria by IPTG induction. We set up two groups of 0 mM IPTG treatment and 0.5 mM IPTG treatment according to the pre-experiment. SDS-PAGE experiments were carried out after low-temperature induction to initially observe the expression of target proteins.
Experiment result
A.Plasmid construction;
B.SDS-PAGE image;
C.SDS-PAGE strip analysis results.
We successfully expressed two proteins, but the amount of expression was not ideal. In the future, we will further explore to optimize the expression and test the engineered bacteria for low temperature resistance.
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 2613
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 2613
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 2404
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 2613
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 2613
Illegal NgoMIV site found at 1366
Illegal NgoMIV site found at 2693 - 1000COMPATIBLE WITH RFC[1000]
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