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:

1. Anaerobic intracellular environment in aerobic extracellular environment which could use to express oxygen sensitive protein
2. most parts in registry optimized for E. coli may also function
3. can use stable genome integration which could transform a larger size of gene at once
4. 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:

1. Nitrogen inducible—repressed by ammonia and other nitrogen source
2. Predicted to express better than the original T7 expression system
3. 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