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Team:Glendale CC AZ/Project
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Contents |
Background
Overview
Growing Desert
Threatened Desert
Wet Lab
Overview
In the past other teams (Osaka, University College London) have explored the resistance genes in the bacteria D. radiodurans. These resistance genes convey resilience against high levels of radiation, oxidative stress and desiccation in that the DNA repairs itself when damaged from these stressors. The interesting thing about these stressors is that the DNA repairs itself in the same way regardless of the type of stress. Simply put, the system responds to DNA damage and makes the necessary repairs. From here, the project first aims to provide extra validation data to those studies. Additionally the ultimate purpose of this iGEM project is to explore the resistance genes in a similar bacteria, Deinococcus hopiensis, as a novel source of these genes within the Deinococcus genus. For this purpose the project then uses the genes to transform a desiccation, radiation, and oxidative stress sensitive bacteria, E. coli to improve the robustness of the cells against the particular stressors of desiccation and oxidative stress. In science, adding validation data is always a very important and welcome resource. In addition to the validation data, the main goal of the project is to look for these same genes in a sister bacteria and see if those genes convey the same resistance.
The Parts
PprI
In previous studies, it was noticed that Osaka had success in decreasing Ionization Radiation in E. coli by transforming the cell with the PprI gene isolated from D. radiodurans. For this group's project, it was decided that the same gene, PprI, would be tested to see if it also increased desiccation resistance in E. coli. To begin the project, the PprI gene that Osaka used was obtained from the IGEM parts registry. This part is identified as part BBa_K602005 in the IGEM parts registry database. When the part was recieved, it was in the form of an agar stab. To isolate the plasmid from the cells in the stab, a miniprep on some of the cells was performed. The product from that miniprep was run on a gel and a band was observed that was in the same size range as the PprI plasmid(insert gel label here). After isolating the PprI plasmid, growth curves using the cells from the agar stab were performed. In these experiments, Rec A, registry part BBa K602003, was used as a control. This is because the Rec A cells had only the coding sequence, no promoter, so Rec A would not have been expressed.
