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Team:DTU-Denmark/Technical stuff lab
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== Lab == | == Lab == | ||
| - | + | \section{Overview} | |
| - | Our project is a proof of concept project, showing that the E. coli sRNA sroB and the intergenic sRNA in the chbBCARG gene can be used to control gene expression by targeting the ybfM Shine-Delgarno, and that these sRNAs can be rationally designed to targeted other Shine-Delgarnos | + | 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 |
| + | |||
The experimental part of the project can be broken into 3 distinct parts, which combine to form the complete project: | The experimental part of the project can be broken into 3 distinct parts, which combine to form the complete project: | ||
| - | + | \begin{itemize} | |
| - | Construction of plasmids necessary for testing our system involves taking the native system from 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. | + | \item Construction of plasmids |
| - | Strain construction involves deleting the original genes from the | + | \item Strain construction |
| - | Improving the araBAD promoter 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 araBAD promoter is used in our project improving this promoter could lead to even finer control of our system. | + | \item Improving the \textit{araBAD} promoter |
| + | \end{itemize} | ||
| + | \paragraph{Construction of plasmids} | ||
| + | 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. | ||
| + | \paragraph{Strain construction} | ||
| + | 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. | ||
| + | \paragraph{Improving the \textit{araBAD} promoter} | ||
| + | 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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Revision as of 15:22, 6 September 2011
Methods
Lab
\section{Overview} 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
The experimental part of the project can be broken into 3 distinct parts, which combine to form the complete project: \begin{itemize} \item Construction of plasmids \item Strain construction \item Improving the \textit{araBAD} promoter \end{itemize} \paragraph{Construction of plasmids} 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. \paragraph{Strain construction} 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. \paragraph{Improving the \textit{araBAD} promoter} 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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Sponsors
Thanks to:
