Team:DTU-Denmark/Technical stuff lab

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{{:Team:DTU-Denmark/Templates/Standard_page_begin|Methods}}
 
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== Lab ==
 
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\section{Overview}
 
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Our project is a proof of concept project, showing that the \textit{E. coli} sRNA \textit{sroB} and the intergenic sRNA in the \textit{chbBCARG} gene can be used to control gene expression by targeting the \textit{ybfM} Shine-Delgarno, and that these sRNAs can be rationally designed to targeted other Shine-Delgarnos
 
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The experimental part of the project can be broken into 3 distinct parts, which combine to form the complete project:
 
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\begin{itemize}
 
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\item Construction of plasmids
 
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\item Strain construction
 
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\item Improving the \textit{araBAD} promoter
 
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\end{itemize}
 
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\paragraph{Construction of plasmids}
 
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necessary for testing our system involves taking the native system from \textit{E. coli}, as well as a slightly modified system and putting them on plasmids that let us both control the expression of these components and measure the output of the system.
 
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\paragraph{Strain construction}
 
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involves deleting the original genes from the chromosome of a \textit{E. coli} W3110 strain. Since we use the original genes, these need to be deleted from the chromosome to prevent them from interfering with our measurements.
 
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\paragraph{Improving the \textit{araBAD} promoter}
 
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entails expanding the dynamic range of this promoter by modifying the -10 and -35 sequence of the promoter, as well as randomly changing the nucleotide sequence around and in between these sequences. Since the \textit{araBAD} promoter is used in our project improving this promoter could lead to even finer control of our system.
 
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Latest revision as of 19:04, 21 September 2011