Team:Calgary/Composite Part
Parts
We submitted three parts to the Registry this year: two parts involved in PHB synthesis and one part involved in PHB secretion.
To meet our medal requirements, we submitted BBa_K2260001 as a new part, which uniquely contains the phaJ4 gene native to Pseudomonas putida, along with the phaC1 gene from Pseudomonas aeruginosa.
We also improved two parts. The first, BBa_K2260000, was an improvement on Imperial College's phaCAB operon submitted in 2013. The gene order was changed to improve PHB yield (Hiroe et al., 2012), codons were optimized for expression in Escherichia coli, and restriction sites were removed to allow part compatibility in all iGEM RFC assembly standards. The second, BBa_K2260002, was an improvement on SDU Denmark's phasin-HlyA part submitted in 2016. Codons were optimized for expression in E. coli.
Synthesis
| Biobrick: BBa_K2260000 |
 |
|---|---|
| Part type: Composite | |
| Description: phaCBA operon with BBa_B0034 RBS | |
| Source: Ralstonia eutropha H16 |
| Biobrick: BBa_K2260001 |  |
|---|---|
| Part type: Composite | |
| Description: phaC1-J4 operon with BBa_B0034 RBS | |
| Source: phaC1 was taken from P. aeruginosa |
Secretion
| Biobrick: BBa_K2260002 |  |
|---|---|
| Part type: Composite | |
| Description: Phasin-HlyA fusion protein with T7 promoter and BBa_B0034 RBS | |
| Source: Phasin (PhaP) is from R. eutropha, HlyA tag is from E. coli, while the T7 promoter is from the T7 bacteriophage. |
Works Cited
Hiroe, A., Tsuge, K., Nomura, C.T., Itaya, M. & Tsuge, T. (2012). Rearrangement of Gene Order in the phaCABOperon Leads to Effective Production of Ultrahigh-Molecular-Weight Poly[(R)-3-Hydroxybutyrate] in Genetically Engineered Escherichia coli. Appl. Environ. Microbiol., 78(9): 3177-3184
