Overview
Parts play the core role in iGEM project and connect us to past or future iGEM team.
We use three kinds of xylitol system in order to make Xylego and each of the main components was used.
The first is a system that absorbs and metabolizes xylitol, feeding it into the pentose phosphate pathway. The second is a system for producing Epidermal Growth Factor (EGF). The third is an RA system for a Kill Switch, which utilizes a tetrathionate-sensitive promoter.
Parts
| Part Registry ID | Part Name | Type | Description |
|---|---|---|---|
| BBa_25M7IBNQ | xylB | Basic | xylB is D-xylulokinase. It changes D-xylulose into D-xylulose 5-phosphate. [1] |
| BBa_25HAUYQA | xdh | Basic | xdh is xylitol dehydrogenase. It changes xylitol into D-xylulose. [2] |
| BBa_2506H3B4 | noxE | Basic | noxE is water-forming NADH oxidase expression gene. |
| BBa_25LO1XY1 | xytB-xytC | Composite | This includes xytB and xytC that are components of the xylitol transporter with xytD BBa_25MSD2OH and xytE BBa_25DHV4J1 . |
| BBa_25MSD2OH | xytD | Basic | This is a component of the xylitol transporter with xytB, xytC BBa_25LO1XY1 and xytE BBa_25DHV4J1 . |
| BBa_25DHV4J1 | xytE | Basic | This is a component of the xylitol transporter with xytB, xytC BBa_25LO1XY1 and xytD BBa_25MSD2OH . |
| BBa_252W5M6H | pBl-xylB-xdh-noxE(T7) | Composite | A pBluescript II SK(-) plasmid carrying xylB BBa_25M7IBNQ , xdh BBa_25HAUYQA , and noxE. |
| BBa_25M65G69 | pTf16-xytB-xytC-xytD-xytE | Composite | A pTf16 plasmid carrying xytB-xytC BBa_25LO1XY1 , xytD BBa_25MSD2OH , and xytE BBa_25DHV4J1 . |
| BBa_254VVDN9 | PrtD-J23118 | Composite | This construct includes the J23118 Anderson promoter and PrtD, which forms a transporter along with PrtE BBa_255K2Q77 and PrtF BBa_2505RRXO . |
| BBa_25WT3TMH | PrtD-lac | Composite | This construct includes the lactose promoter and PrtD, which forms a transporter along with PrtE BBa_255K2Q77 and PrtF BBa_2505RRXO . |
| BBa_255K2Q77 | PrtE | Basic | This is a component of the transporter with PrtD and PrtF BBa_2505RRXO . |
| BBa_2505RRXO | PrtF | Basic | This is a component of the transporter with PrtD and PrtE BBa_255K2Q77 . |
| BBa_25IXDM9P | EGF-Lard3 | Composite | By fusing the Lipase ABC transporter Recognition Domain 3 (LARD3) to the C-terminus of Epidermal Growth Factor (EGF), we enable its extracellular transport through the PrtD, PrtE BBa_255K2Q77 , and PrtF BBa_2505RRXO pathway. |
| BBa_254BZVQT | pTf16-lac-PrtD-PrtE-PrtF | Composite | PrtD-lac BBa_25WT3TMH , PrtE BBa_255K2Q77 , and PrtF BBa_2505RRXO were inserted into pTf16. |
| BBa_25FLJF7S | pTf16-J23118-PrtD-PtrE-PtrF | Composite | J23118-PrtD BBa_254VVDN9 , PrtE BBa_255K2Q77 , and PrtF BBa_2505RRXO were inserted into pTf16. |
| BBa_25SLGMUS | 6×His-EGF-Lard3 | Composite | A 6×His sequence was added to the N-terminus of EGF-LARD3. This facilitates purification and allows the use of versatile anti-6×His antibodies. |
| BBa_25SA83H1 | pBl-EGF-Lard3 | Composite | EGF-LARD3 BBa_25IXDM9P was inserted under the T7 promoter of pBluescript II SK(-). |
| BBa_25IRUXQA | pBl-6×His-EGF-Lard3 | Composite | 6×His-EGF-LARD3 BBa_25SLGMUS was inserted under the T7 promoter of pBluescript II SK(-). |
| BBa_25HGCNHF | TtrS | Basic | It phosphorylates TtrR BBa_K3972001 in response to tetrathionate and activates the promoter downstream of TtrR. [3] |
| BBa_25RY44EP | pTf16-TtrS | Composite | TtrS BBa_25HGCNHF was inserted under the J23118 Anderson promoter of pTf16. |
| BBa_K3972001 | TtrR | Basic | It is phosphorylated by TtrS in response to tetrathionate and activates the promoter downstream of TtrR. [3] |
| BBa_250GIM06 | BsaIM1 | Basic | It is a restriction-modification enzyme that methylates DNA. [4] |
| BBa_253BD4YH | BsaIM2 | Basic | It is a restriction-modification enzyme that methylates DNA. [4] |
| BBa_25A1HE90 | BsaIR | Basic | It is a restriction enzyme with a cleavage site in the ampicillin resistance gene. [4] It is more stable than BsaIM. |
References
- Wungsintaweekul J, Herz S, Hecht S, Eisenreich W, Feicht R, Rohdich F, et al. Phosphorylation of 1‐deoxy‐D‐xylulose by D‐xylulokinase of Escherichia coli. European Journal of Biochemistry. 2001 Jan;268(2):310–6.
- Zhou P, Li S, Xu H, Feng X, Ouyang P. Construction and co-expression of plasmid encoding xylitol dehydrogenase and a cofactor regeneration enzyme for the production of xylitol from d-arabitol. Enzyme and Microbial Technology. 2012 July;51(2):119–24.
- Daeffler KN, Galley JD, Sheth RU, Ortiz‐Velez LC, Bibb CO, Shroyer NF, et al. Engineering bacterial thiosulfate and tetrathionate sensors for detecting gut inflammation. Mol Syst Biol. 2017 Apr 3;13(4):923.
- Geobacillus stearothermophilus M1.BsaI (bsaIM1), M2.BsaI (bsaIM2), and BsaI (bsaIR) genes, complete cds [Internet]. 2006 [cited 2025 Oct 7]. Available from: http://www.ncbi.nlm.nih.gov/nuccore/AY453694.1