IGEM:UNAM Genomics Mexico/2009/Notebook/Wifi coli/2010/06/24

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News 1. The 2010 plate will arrive within the next days. 2. There will be a congress in Irapuato on July 5, 2010. Whoever wants to go is welcome, though trip expenses are on us. 3. There are new deadlines in the openwetware. We MUST have our proyect explained broadly, a project name, and the corresponding logo by July 16, 2010. As most of us are going on holidays, it is extremely important to have things done ahead so that nothing is left unattended. 4. Jose Luis will help us to use special equipment in the Institute of Biotechnology (IBT), UNAM, to make assays of absorbance and luminescence. Wet lab This week, we were received EnvZ strain. Augusto has already plated it and bacteria is growing out well. Mariana is working in the punctual mutation of luciferase. By the time that it is ready, she will ligate it into a plasmid with a constitutive promoter, transform it and to check that the emission spectrum presents a change, she will plan a experiment involving the usage of the special equipment in the IBT. We finnaly could insert the minimum blue promoter into plasmid 30 pSB4A5 by PCR. This PCR product will be purified and GFP reporter will be inserted into this PCR product. As GFP reporters we’ll use GFP E0240, recommended in the BioBrick Promoter Measurement Kit, and also GFP BBa_K145015, this reporter is well characterized by the KU Leuven 2008 team and has a half-life of 74 minutes. To test Blue Promoter functionality, Jorge will use the iGEM BioBrick Promoter Measurement Kit. For this test he has done transformations, plasmid extraction and restrictions to the GFP reporter (E0240), a backbone plasmid (pSB3K3) and BBa_I20260, a GFP regulated under the constitutive promoter J23101, as reference promoter. Ligation with Blue Promoter + GFP reporters (the two GFPs described above) + Backbone plasmid will be performed by three different ways: 1. First ligate Blue Promoter + GFP reporters, and then ligate this construct to the backbone plasmid. 2. Blue Promoter + GFP reporters + Backbone plasmid by a 3-way ligation. 3. Insert the Minimum Blue Promoter to backbone plasmid by PCR and then ligate GFP reporters to this PCR product. Jorge finally has done the transformations with a YFP reporter BBa_K117008 that will be used for the fluorescence assay. Modeling This week: OmpF-OmpC experiment desing in order to use OmpFp as a promoter for LuxAB. Simbiology modeling for the different systems. Daniela and Claudia are working in the Lux Operon modeling in simbiology. Daniela is also in charge of the oligos' design for the OmpFp. Hector and Amhed will be working on the assay to test the blue promoter (along with Jorge's work).