Team:NAU-CHINA/PARTS
PARTS
Overview
This year, NAU-CHINA has designed a total of 19 parts, including 2 new basic parts and 17 composite parts.
We designed BBa_K3735001(sefA) to enhance the stability of SeNPs and the tolerance of our engineered bacteria to selenite, and BBa_K3735002(ssuE) to improve the reduction efficiency of selenite for E.coli BL21(DE3).
BBa_J04450 is the most prevalent part for signal detection. But it may not be reliable any more when we conduct lower expression experiments. To solve this problem, this year, we improved BBa_J04450 to make it stronger so that we can detect it easier in the same condition.
If you want to see more details, you can find the usage and characterization in the part registry page. You can also visit the basic part, composite part, part collection and improved part page for more information.
| Part Number | Name | Type |
|---|---|---|
| BBa_K3735001 | SefA | Coding |
| BBa_K3735002 | SsuE | Coding |
| BBa_K3735003 | J23105-B0034-K3735001 | Composite |
| BBa_K3735004 | J23101-B0034-K3735001 | Composite |
| BBa_K3735005 | J23119-B0034-K3735001 | Composite |
| BBa_K3735006 | J23101-B0034-K3735002 | Composite |
| BBa_K3735007 | J23105-B0034-K3735002 | Composite |
| BBa_K3735008 | J23119-B0034-K3735002 | Composite |
| BBa_K3735009 | J23119-B0034-K3735001-J23119-B0034-K3735002 | Composite |
| BBa_K3735010 | J23119-B0034-K3735001-J23101-B0034-K3735002 | Composite |
| BBa_K3735011 | J23119-B0034-K3735001-J23105-B0034-K3735002 | Composite |
| BBa_K3735012 | J23101-B0034-K3735001-J23119-B0034-K3735002 | Composite |
| BBa_K3735013 | J23101-B0034-K3735001-J23101-B0034-K3735002 | Composite |
| BBa_K3735014 | J23101-B0034-K3735001-J23105-B0034-K3735002 | Composite |
| BBa_K3735015 | J23105-B0034-K3735001-J23119-B0034-K3735002 | Composite |
| BBa_K3735016 | J23105-B0034-K3735001-J23101-B0034-K3735002 | Composite |
| BBa_K3735888 | J23105-B0034-K3735001-J23105-B0034-K3735002 | Composite |
| BBa_K3735018 | 5'UTR-E1010 | Coding |
| BBa_B0034 | RBS | RBS |
| BBa_J23101 | Middle Promoter | Promoter |
| BBa_J23105 | Low Promoter | Promoter |
| BBa_J23119 | High Promoter | Promoter |
| BBa_K608011 | Contribution | Composite |
| BBa_J04450 | R0010-B0034-E1010-B0015 | Composite |
| BBa_K3735666 | improvement | Composite |
| BBa_K3735020 | improvement | 5'UTR |
Basic Parts
NAU-CHINA 2021 designed and used two new basic parts: BBa_K3735001 sefA coding part and BBa_K3735002 ssuE coding part, aiming to increase the efficiency and quality of producing SeNPs.
BBa_K3735001 is used to express SefA protein and wrap the elemental selenium, in order to increase the tolerance to selenium pollution and produce SeNPs with high biological activity.
BBa_K3735002 is used to express NAD(P)H dependent flavin oxidoreductase(ssuE), enhancing the biological reduction ability of our engineered bacteria.
You can click on the part number for more details in the following table.
| Part Number | Name | Type |
|---|---|---|
| BBa_K3735001 | SefA | Coding |
| BBa_K3735002 | ssuE | Coding |
| BBa_B0034 | RBS | RBS |
| BBa_J23101 | Middle Promoter | Promoter |
| BBa_J23105 | Low Promoter | Promoter |
| BBa_J23119 | High Promoter | Promoter |
| BBa_K3735020 | improvement | 5'UTR |
Composite Parts
To reach the optimal balance between metabolic burden and productivity, we designed 15 composite components by adjusting promoter to find the most efficient combination.
We first verified the feasibility of each part separately.
Then we combined sefA and ssuE with different promoters and find the best expression intensity to produce SeNPs by analyzing growth indicators and the final production efficiency.
| Part Number | Name | Type |
|---|---|---|
| BBa_K3735003 | J23105-B0034-K3735001 | Composite |
| BBa_K3735004 | J23101-B0034-K3735001 | Composite |
| BBa_K3735005 | J23119-B0034-K3735001 | Composite |
| BBa_K3735006 | J23101-B0034-K3735002 | Composite |
| BBa_K3735007 | J23105-B0034-K3735002 | Composite |
| BBa_K3735008 | J23119-B0034-K3735002 | Composite |
| BBa_K3735009 | J23119-B0034-K3735001-J23119-B0034-K3735002 | Composite |
| BBa_K3735010 | J23119-B0034-K3735001-J23101-B0034-K3735002 | Composite |
| BBa_K3735011 | J23119-B0034-K3735001-J23105-B0034-K3735002 | Composite |
| BBa_K3735012 | J23101-B0034-K3735001-J23119-B0034-K3735002 | Composite |
| BBa_K3735013 | J23101-B0034-K3735001-J23101-B0034-K3735002 | Composite |
| BBa_K3735014 | J23101-B0034-K3735001-J23105-B0034-K3735002 | Composite |
| BBa_K3735015 | J23105-B0034-K3735001-J23119-B0034-K3735002 | Composite |
| BBa_K3735016 | J23105-B0034-K3735001-J23101-B0034-K3735002 | Composite |
| BBa_K3735888 | J23105-B0034-K3735001-J23105-B0034-K3735002 | Composite |
| BBa_K3735018 | 5'UTR-E1010 | Coding |
| BBa_K608011 | J23110-B0032-E0040 | Composite |
| BBa_J04450 | R0010-B0034-E1010-B0015 | Composite |
| BBa_K3735666 | R0010-B0034-K3735018-B0015 | Composite |
Parts Collection
This year NAU-CHINA team combined 2 genes(sefA (BBa_K3735001) and ssuE (BBa_K3735002)) with optimal promoters as a collection.
Selenite is a toxic substance to E.coli while SefA is an important protein which can wrap elemental selenium and increased bacteria’s tolerance to selenite. ssuE may help enhance the bacteria's bio-reduction,also help bacteria survive in selenite pressure.
At the beginning we designed nine expression combinations but after mathematical modeling we chose the low intensity sefA and low intensity ssuE as our gene pathway which is the optimal circuit for SeNPs production. (BBa_K3735013, see results for details)
| Part Number | Name | Type |
|---|---|---|
| BBa_K3735888 | J23101-B0034-K3735001-J23101-B0034-K3735002 | Composite |
| BBa_K3735003 | J23105-B0034-K3735001 | Composite |
| BBa_K3735007 | J23105-B0034-K3735002 | Composite |