Team:Utrecht/Parts
Several biobricks from the distribution kit were used during our project. These were retrieved from the kit using the protocols provided by iGEM, transformed into E. coli, isolated in elution buffer NE. We also ordered several constructs from IDT (Table 1). These Biobricks were used to create several new biobricks that were relevant to our project. BBa_K2736000 and BBa_K2736001, were created to be used as BRET pair to measure the activity of our customized Tar receptors BBa_K2736003, BBa_K2736004, and BBa_K2736005. To test whether it was possible to switch ligand binding domains of the Tar receptor and two component receptors we created BBa_K2736002, and BBa_K2736006.
Used Biobricks
| Name | Biobrick Number | Size | Size | Obtained |
|---|---|---|---|---|
| RFP | BBa_K516032 | 828 | Red fluorescence | isolated |
| Promoter | BBa_K608003 | 56 | Strong promoter with medium ribosome binding site | isolated |
| eYFP | BBa_I15017 | 724 | Yellow fluorescence | isolated |
| CheZ | BBa_K629003 | 645 | "cheZ chemotaxis regulator protein phosphatase for CheY" | isolated |
| Copper promoter | 98 | Promoter responsive to phosphorylated CusR | ordered | |
| Custom receptor | BBa_K2736006 | 1505 | Fusion protein of CusS with the Tar ligand binding domain | ordered |
| 1/5A::CheZ::Rluc | BBa_K2736002 | 1655 | CheZ fused to Rluc | ordered |
| CheY | 390 | Chemotaxis response regulator protein | ordered |
Created Biobricks
Here comes Table 2