Impact of 23S rRNA single mutations on growth rates

We developed Statistical Coupling Analysis (SCA) for RNA which predicts allosteric networks, or sectors, in complex RNAs. We applied our analysis to the 23S rRNA as a test system and used it to make predictions about the functional importance of nucleotides in the 23S. To validate our predictions, we performed a continuous culture experiment on E. coli SQZ10 transformed with PLK35 plasmids harboring a 23S rRNA gene with various mutations. We used next generation sequencing to calculate the relative frequency of mutations throughout the continuous culture experiment in order to determine the impact the mutations had on growth rate.