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Team:BIOTEC Dresden/Notebook Vesicles
From 2009.igem.org
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Vesicles are formed using a microfluidic system, allowing for small reaction volumes and a controllable environment. The system is used to create emulsions of material in aqueous phase in a lipid phase. Proteins and various buffers are eventually added to the aqueous phase to carry out transcription and translation of the protein of interest in the vesicles, in this case, FLP recombinase. The production of FLP can be checked by tagging the final protein with a fluorescent protein like GFP, which can be then viewed using fluorescent microscopy. | Vesicles are formed using a microfluidic system, allowing for small reaction volumes and a controllable environment. The system is used to create emulsions of material in aqueous phase in a lipid phase. Proteins and various buffers are eventually added to the aqueous phase to carry out transcription and translation of the protein of interest in the vesicles, in this case, FLP recombinase. The production of FLP can be checked by tagging the final protein with a fluorescent protein like GFP, which can be then viewed using fluorescent microscopy. | ||
Revision as of 23:45, 21 October 2009
Gene Expression in vesicles
The first step was to create a microfluidic chamber that creates vesicles of homogenous size. In the second step, a commercial gene expression kit was introduced into the vesicles and tested using a GPF plasmid.
Strategy
Vesicles are formed using a microfluidic system, allowing for small reaction volumes and a controllable environment. The system is used to create emulsions of material in aqueous phase in a lipid phase. Proteins and various buffers are eventually added to the aqueous phase to carry out transcription and translation of the protein of interest in the vesicles, in this case, FLP recombinase. The production of FLP can be checked by tagging the final protein with a fluorescent protein like GFP, which can be then viewed using fluorescent microscopy.
The protocols used to create the microfluidic chamber and express genes in it are given here.
Following that protocol, chambers were produced and gene expression was attempted. Here are the results obtained during that process.
