Azotobacter vinelandii
Although Azotobacter Vinelandii is not commonly used as a model of gram-negative bacteria, there are many features that make us introduce the chassis to iGEM. We think that it is very suitable for genetic engineering based on the features below:
- Anaerobic intracellular environment in aerobic extracellular environment which could use to express oxygen sensitive protein
- most parts in registry optimized for E. coli may also function
- can use stable genome integration which could transform a larger size of gene at once
- Antibiotic such as ampicillin and kanamycin can be used and iGEM vectors such as PSB1A3 can be used
Nitrogen-repressible T7 Expression System
For introducing the stable genome integration, we constructs several biobricks. Moreover, as many promoter of registry cannot be used in Azotobacter Vinelandii, we introduce a novel T7 dependent system using nifH promote. Features about the protein expression system:- Nitrogen inducible—repressed by ammonia and other nitrogen source
- Predicted to express better than the original T7 expression system
- Make T7 promoter usable in Azotobacter Vinelandii, and hence, make a lot more biobricks could be used in Azotobacter Vinelandii.
You can find the parts constructed for stable integration and T7 expression here: IGEM 2014 CUHK--biobricks
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