Team:TU-Eindhoven/Production

From 2013.igem.org

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These proteins were selected using the RCSB Protein Data Bank. A python program was designed by us alongside a molecular dynamics simulator, to ensure that the selected proteins had a high content of Arginine and Lysine aminoacids. The complete description of the protein selection process can be found on the [[Team:TU-Eindhoven/ProteinSelection| protein selection section of the drylab tab.]] The rest of the proteins were selected using the results of McMahon's research (2008) [1].
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These proteins were selected using the RCSB Protein Data Bank. A python program was designed by us alongside a molecular dynamics simulator, to ensure that the selected proteins had a high content of Arginine and Lysine amino acids. The complete description of the protein selection process can be found on the [[Team:TU-Eindhoven/ProteinSelection| protein selection section of the drylab tab.]] The rest of the proteins were selected using the results of McMahon's research (2008) [1].
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These protein sequences were transformed into pET28a vectors for aerobic expression. When the proteins were overexpressed and when the cell pellets were fixated in 4% PFA (paraformaldehyde) for two days, MRI CEST measurements were performed and the data was processed. For more detailed information about the lab procedures and the steps that were taken to come to these results, see [https://2013.igem.org/Team:TU-Eindhoven/CESTAgentTesting CEST Agent Testing], for the results of the data processing, see [https://2013.igem.org/Team:TU-Eindhoven/MRIProcessing MRI data processing].  
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These protein sequences were transformed into pET28a vectors for aerobic expression. When the proteins were over expressed and when the cell pellets were fixated in 4% PFA (paraformaldehyde) for two days, MRI CEST measurements were performed and the data was processed. For more detailed information about the lab procedures and the steps that were taken to come to these results, see [https://2013.igem.org/Team:TU-Eindhoven/CESTAgentTesting CEST Agent Testing], for the results of the data processing, see [https://2013.igem.org/Team:TU-Eindhoven/MRIProcessing MRI data processing].  
== References ==
== References ==

Revision as of 23:05, 18 October 2013

Potential CEST Contrast Agents

Protein production is induced by the FNR promoter. Each of the 8 proteins listed below can be expressed once a hypoxic environment is sensed. These proteins are all rich in Arginine and Lysine amino acids, which are known to be good CEST candidates.


The proteins that the CEST protein production device can generate are:


These proteins were selected using the RCSB Protein Data Bank. A python program was designed by us alongside a molecular dynamics simulator, to ensure that the selected proteins had a high content of Arginine and Lysine amino acids. The complete description of the protein selection process can be found on the protein selection section of the drylab tab. The rest of the proteins were selected using the results of McMahon's research (2008) [1].

These protein sequences were transformed into pET28a vectors for aerobic expression. When the proteins were over expressed and when the cell pellets were fixated in 4% PFA (paraformaldehyde) for two days, MRI CEST measurements were performed and the data was processed. For more detailed information about the lab procedures and the steps that were taken to come to these results, see CEST Agent Testing, for the results of the data processing, see MRI data processing.

References

[1] McMahon, M.T., 2008, New “Multi-Color” Polypeptide DIACEST Contrast Agents for MR Imaging, Magn Reson Med. 2008 October ; 60(4): 803–812.