Team:Technion-Israel/Basic Part
Basic Parts
Our new basic parts include the Invertase (BBa_K2934001) and the Glucose Oxidase (GOx) (BBa_K2934000) genes, both are major enzymes in the production process of honey in the bees’ stomach.
These enzymes are responsible, respectively, for sucrose degradation to glucose and fructose and the oxidation of glucose to D-glucono-lactone. Both enzymes’ sequences are derived from A. niger and optimized for our model organism, Bacillus subtilis.
The third enzyme that plays an important role in the honey process is catalase, which is a native enzyme to B. subtilis.
In our lab work, we used TaKaRa’s commercial plasmid pBE-S that is designed to secrete recombinant proteins from B. subtilis. Some of our basic parts include several parts from that plasmid, including an RBS and a secretory signal peptide-protein linker.
List of Our Basic Parts
| Name | Type | Description | Designer | Length | |||
|---|---|---|---|---|---|---|---|
| Favorite | W | BBa_K2934000 | Coding | Glucose Oxidase-Histag A. niger optimized for B. subtilis | Nir Litver, Lior Haim | 1857 | |
| Favorite | W | BBa_K2934001 | Coding | Invertase-Histag A. niger optimized for B. subtilis | Nir Litver, Lior Haim | 1809 | |
| BBa_K2934002 | Regulatory | pKatA promoter for Bacillus subtilis | Shira Levi | 66 | |||
| BBa_K2934003 | Terminator | AmyE 3' UTR from Bacillus subtilis | Nir Litver | 75 | |||
| BBa_K2934005 | Protein Domain | Signal peptide-protein linker for Bacillus subtilis | Nir Litver | 27 | |||
| BBa_K2934007 | RBS | 5' UTR derived from pBE-S for Bacillus subtilis | Nir Litver, Shira Levi | 40 |
Our Best New Basic Part
Our best new basic part is BBa_K2934002, the B. subtilis catalase promoter (pKatA).
This promoter is interestingly regulated – its repressor, PerR, binds iron and zinc ions and can only bind the promoter in absence of hydrogen peroxide. Hydrogen peroxide presence leads to Fe2+-catalyzed oxidation of histidines, which interferes with the repressor’s ability to bind the promoter and resulting in the expression of the PerR-regulated gene[1]. Thus, the downstream gene can only be transcribed in the presence of hydrogen peroxide.
We believe that this part can be useful for future iGEM teams who wish to express genes in a controlled manner in Bacillus subtilis.
Figure 1: The effect of hydrogen peroxide on PerR and its affinity to the pKatA promoter.
- Lee JW, Helmann JD. 2006. The PerR transcription factor senses H2O2 by metal-catalysed histidine oxidation. Nature 440:363–367.
