Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Luna Barroco de Lacerda
Date created: 2014-10-05 11:00:00
Date modified: 2015-05-08 01:10:02
Mercury ions detector device
| Types | DnaRegion |
| Roles | engineered_region Device |
| Sequences | BBa_K1355002_sequence (Version 1) |
Description
We used the biobrick (BBa_E0840) available in the database of the iGEM since 2004 to be connected to our ???essential biobrick??? (BBa_K1355001) and - together - form a system capable of biodetect mercury in bacteria.In the presence of mercury, The MerR protein will bind to it and MerT, MerP and the Green Fluorescent Protein (GFP) protein will be expressed and the biodetection will begin! The MerP and MerT protein are responsible for the transport of mercury from the periplasm to the cytoplasm and the GFP will act as rep??rter to alerting the presence of mercury in the middle and the higher concentration of mercury is, higher will be the GFP expression. And If there is no mercury in the middle, the synthesis of MerT, MerP and GFP protein will be repressed by the MerR repressor protein.
Notes
For this construct, we followed these steps: Extraction of plasmid DNA and DNA quantification of bacteria transformed with the Essential Biobrick (BBa_K1355001) and the bacteria transformed with the GFP Biobrick (BBa_E0840); Verifying the electrophoretic profile of the extracted plasmid DNA; Restriction enzyme digestion for Essential Biobrick with SpeI and PstI and for GFP Biobrick with PstI and XbaI; Checking the electrophoretic profile of digested samples; Fragment of interest purification from the gel; Ligation the gel purification of GFP gene and digested Essential Biobrick; Transformation of ligation in DH5-alpha; Plasmid DNA extraction from bacteria transformed; Check the electrophoretic profile to see results of samples linked (no fragments); Digestion of Essential Biobrick + GFP Biobrick with EcoRI and PstI, aiming to analyze the fragment size to be isolated; Checking the electrophoretic profile of the digested sample to obtain results showing that the isolated fragment is the junction of Essential Biobrick +GFP Biobrick ;Source
BBa_K1355001, BBa_E0840| Access | Instance | Definition |
| public public | BBa_K1355001 BBa_E0840 | BBa_K1355001 GFP genera |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_K1355001 BBa_E0840 | 1,1189 1198,2075 | BBa_K1355001 GFP genera |