All About Parts We Created
The gene for PCB detection was a modified luciferase gene. The genes for PCB degradation were based on those used by 2017 iTelsa SoundBio team and described in this the 2019 Ewald et al. paper . The promoter for Rhodococcus jostii is one that was published in Round et al.
| Name | Type | Description | Length |
|---|---|---|---|
| BBa_K4458001 | Basic part | pcbA1 gene from Dehalococcoides mccartyi | 1512 bp |
| BBa_K4458003 | Basic part | pcbA4 from Dehalococcoides mccartyi | 1407 bp |
| BBa_K4458000 | Basic part | pcbA5 from Dehalococcoides mccartyi | 1446 bp |
| BBa_K4458002 | Basic part | PM6-OP3 promoter for Rhodococcus | 250 bp |
| BBa_K4458004 | Composite part | pcbA1 expression construct | 1925 bp |
| BBa_K4458005 | Composite part | pcbA4 expression construct | 1820 bp |
| BBa_K4458006 | Composite part | pcbA5 expression construct | 1859 bp |
| BBa_K4458010 | Composite part | CYP1A1-CYC1-Akaluc | 139 bp |
| BBa_K4458011 | Basic part | Aryl Hydrocarbon Receptor Nuclear Translocator (ARNT) | 2547 bp |
| BBa_K4458012 | Basic part | Aryl Hydrocarbon Receptor (AHR) | 2547 bp |
| BBa_K4458013 | Basic part | Aryl Hydrocarbon Receptor Nuclear Translocator (ARNT) | 2325 bp |
| BBa_K4458014 | Basic part | Cytochrome P450 Family 1 Subfamily A Member 1 (CYP1A1) | 3572 bp |
| BBa_K4458015 | Basic part | Akaluc | 1653 bp |
| BBa_K4458016 | Composite part | PCB Biosensor | 5600 bp |
| BBa_K4458017 | Composite part | PCB Biosensor | 5578 bp |
| BBa_K4458018 | Composite part | AHR Expression System | 5558 bp |
Akaluc, or Akalumine-Luciferase, is a modified luciferase complex designed to increase expression when exposed to PCBs. Luciferase is a naturally occurring bioluminescent protein found in species like fireflies which would allow our bioreactor to glow.
pcbA1, like all the other genes in this set, are dehalogenases extracted from Dehalococcoides species, meaning they remove halogens such as chlorine from molecules. pcbA1 specializes in the removal of meta halogens, which are halogens neither adjacent nor opposite to the primary carbon in a ring.
pcbA4 is similar to pcbA1 in that it is a dehalogenase. However, this enzyme is specialized in the removal of para halogens, which are opposite the primary carbon. Also, pcbA4 accumulates in cells faster than either of its related enzymes, according to the 2019 Ewald et al. paper
pcbA5 is also a dehalogenase. However, it can remove para and meta chlorines. Notably, none of these enzymes remove ortho chlorines, which are right next to the central carbon. If we continued this project, we could try to design an enzyme that targets those ortho chlorines.
This promoter was identified in the paper Round et. al as the most active promoter for Rhodococcus. We used this and added it to the registry because of the lack of other information about Rhodococcus species in iGEM.