Team:HFLS ZhejiangUnited/Parts
Summary
Our project is aim to construct a plasmid with formaldehyde detection, color display, and formaldehyde degradation on an existing basis. There are two strategies for formaldehyde detection (HxlR and frmR), and also two strategies for formaldehyde degradation (PADH+FDH and FDM). Therefore, there are four possible choices for the combinations.
1. BBa_K2890006: HxlR + eRFP + FDM
The sequence is composed of formaldehyde promoter HxlR (BBa_K1334002), gene RFP, and formaldehyde degrading gene FDM (optimized FDM gene BBa_K2890003).
When the pathway works, we can see the E.coli turns to red with naked eyes at the presence of formaldehyde. Then FDM gene will be further expressed to biodegrade formaldehyde.
2. BBa_K2890005: HxlR+PADH+FDH
The sequence is composed of formaldehyde promoter HxlR (BBa_K1334002), gene RFP, and formaldehyde degrading gene PADH+FDH (BBa_K2890002).
3. BBa_K2890007: frmR+eRFP+PADH+FDH
The sequence is composed of formaldehyde promoter frmR (BBa_K2890001), eRFP (BBa_K1357010) and PADH+FDH (BBa_K2890002).
This part was successfully constructed, but did not work as we expected.
4. BBa_K2890008: frmR+eRFP+ FDM
The sequence is composed of formaldehyde promoter frmR (BBa_K2890001), eRFP (BBa_K1357010) and FDM (BBa_K2890003).
This part was successfully constructed, but did not work as we expected.
5. BBa_K2890001: optimized frmR
An optimized frmR promoter for formaldehyde detection.
We optimized the coding sequence from the original part (BBa_K749004), and add a pFrmAB_RBS at the end.
6. BBa_K2890002: optimized PADH + FDH
A optimized combined PADH plus FDH used for degredation of formaldehyde.
The coding regions were optimized for more effective expression of both genes in E. coli. For further increasing the expression, promoters were added for both genes.
Optimized and modified from BBa_K1706002 and BBa_K1706005, by adding separate promoters to upstreams of each coding sequences.
7. BBa_K2890003: optimized FDM
Optimized and modified FDM gene based on the original part BBa_K2310900. FDM was used for biodegrading formaldehyde, by converting it to methanol.
8. BBa_K2890004: HxlR+tsPurple
This part is combined and modified with standard BioBricks parts BBa_K1334002 and BBa_K1357008.
A HxlR-based formaldehyde detecting reporter part, displaying by purple color.
9. BBa_K2890011: HxlR+amilCP
This part is combined BBa_K1334002 (HxlR) with BBa_K1357009 (amilCP).
A HxlR-based formaldehyde detecting reporter part, displaying by blue color.
Parts
Figure 1 Composition of part BBa_K2890006.
Figure 2 Composition of part BBa_K2890005.
Figure 3 Composition of part BBa_K2890007.
Figure 4 Composition of part BBa_K2890008.