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.

Bla..