PRJNA528665
Molecular evolution of TEM-1 beta lactamase by epPCR
This archive contain the TEM-1 beta lactamase datasets used for the validation of CAMELS (Coupling Analysis by Molecular Evolution Library Sequencing). This dataset were used to prove the possibility to use Direct Coupling Analysis (DCA) to retrieve structural information from molecular evolution libraries. DCA is a powerful technique that enables to extract structural information of proteins belonging to large protein families exclusively by in silico analysis. This method is however limited by sequence availability and various biases. We proposed a new method (CAMELS) that exploits molecular evolution to circumvent these limitations: instead of relying on existing protein families, we used in vitro mutagenesis of TEM-1 beta lactamase combined with in vivo functional selection to generate the sequence data necessary for evolutionary analysis. We could reconstruct by this strategy the lactamase fold exclusively from sequence data. Through generating and sequencing large libraries of variants, it is possible to deal with any protein, ancient or recent, from any species, having the only constraint of setting up a functional phenotypic selection of the protein. This method allows us to obtain protein structures without solving the structure experimentally.