Team:Utrecht/Parts
Several biobricks from the kit were used during our project. Some of these were isolated from the kit by direct transformation of 2 μl of DNA resuspended in elution buffer NE. Besides the biobricks that were isolated from the kit, some constructs were ordered completely 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 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 | 1655CheZ fused to Rluc | ordered | |
| CheY | 390 | Chemotaxis response regulator protein | ordered |
Created Biobricks
Here comes Table 2