Team:Heidelberg/Project Highlights
From 2009.igem.org
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Applying both qRT-PCR and flow cytometry, we developed two new relative units in analogy to bacterial RPU for use in mammalians: Relative Expression Units and Relativeb Mammalian Promoter Units. We apply these units to the characterization of our promoters, as well as an existing [http://partsregistry.org/Part:BBa_I712004 promoter from the registry]. | Applying both qRT-PCR and flow cytometry, we developed two new relative units in analogy to bacterial RPU for use in mammalians: Relative Expression Units and Relativeb Mammalian Promoter Units. We apply these units to the characterization of our promoters, as well as an existing [http://partsregistry.org/Part:BBa_I712004 promoter from the registry]. | ||
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+ | [[Image:HD09_CMV_standard3.png|center|400px|thumb|'''Strength of the CMV promoter in different cell lines in REU'''. [https://2009.igem.org/Team:Heidelberg/Project_Measurement Relative Expression Units (REU)] was determined by Flow cytometry; in HeLa and MCF-7 cells, this measurement was confirmed by an independent technique (Image analysis). 3 experiments on different days, three replicates for each measurement.]] | ||
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+ | [[Image:Qpcr_result.png|center|400px|thumb|'''Real-time RT-PCR data of cmv promoter'''. One group of HeLa cells were transfected with plasmid containing CMV promoter coupled to GFP. Another group was transfected with JeT promoter coupled to GFP served as reference. Total RNA generated by CMV was divided by total RNA generated by JET to obtain [https://2009.igem.org/Team:Heidelberg/Project_Measurement RMPU] .]] | ||
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== 4 RFCs, well characterized parts == | == 4 RFCs, well characterized parts == |
Revision as of 16:13, 21 October 2009
Project HighlightsWe are able to predict functional mammalian promoter sequencesWe created HEARTBEAT, a model portfolio based upon genome-wide bioinformatic analysis and fuzzy logic. We used these tools to predict promoter sequences. Then, we synthesized such sequences. We found that promoters most closely matching the model predictions work, whereas others don't. Read more about these results on the "HEARTBEAT database" page. We also present a GUI which enables you to design the promoter of your needs. We have created a functional biochemical synthesis method for the generation of promoters librariesWe developed RA-PCR, a biochemical method for the generation of randomized promoter libraries. We created a library of constitutive promoters, a NF-κB responsive promoter which underwent extensive characterization and some other regulated promoters. We have developed novel standards for measurement of promoters in mammalian cellsApplying both qRT-PCR and flow cytometry, we developed two new relative units in analogy to bacterial RPU for use in mammalians: Relative Expression Units and Relativeb Mammalian Promoter Units. We apply these units to the characterization of our promoters, as well as an existing [http://partsregistry.org/Part:BBa_I712004 promoter from the registry]. 4 RFCs, well characterized partsWe stick with the standards required by synthetic biology and submit all the methods and units we developed as RFCs (Request for Comments):
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