Hello all.
I am attempting to generate a bacterial strain that expresses two separate fluorescent proteins by placing the proteins into a single plasmid. To do so I plan on cloning the one fluorescent protein including its promoter into a plasmid already containing the separate fluorescent protein and promoter.
I would like to know what the factors I would need to consider are, when choosing the two promoters for this system, to ensure that they are compatible. Any assistance will be much appreciated.
Thank you.
These papers might help you.
https://www.hindawi.com/journals/isrn/2013/590587/
Article A set of dual promoter vectors for high throughput cloning, ...
https://academic.oup.com/femsle/article/232/2/173/536508
Article Two-promoter vector is highly efficient for overproduction o...
There should not be any compatibility issues with having two different promoters on the same plasmid. If you think about it, plasmids normally carry multiple promoters anyway, one for the drug resistance gene, frequently one or two for plasmid replication, and then whatever promoter might be added to the plasmid. So you can pick whichever pair of promoters you might like. Just be sure that you have figured out appropriate regulation of the promoters and whether you also need to include regulatory genes on the plasmids or not. Ideally you would have a good transcription terminator separating the two so that transcription from the upstream one does not also transcribe your downstream gene. Or you could have them pointing in opposite directions.
- Badges
- Science topic