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Mapping of Amino Acid Substitutions Conferring Herbicide Resistance in Wheat Glutathione Transferase.

Govindarajan, Mannervik, Silverman, Wright, Regitsky, Hegazy, Purcell, Welch, Minshull, Gustafsson.
ATUM (DNA2.0), Calysta, National Research Center Egypt, Stockholm University
ACS Synthetic Biology 2015
Gene names: Glutathione transferase (GST). Host systems: Escherichia coli BL21[Bacterial]. Gene species: Triticum aestivum[Plant]. Optimized: Yes. Protein activity: Yes. Vectors: pJ401. Tags: His6.
Abstract: We have used design of experiments (DOE) and systematic variance to efficiently explore glutathione transferase substrate specificities caused by amino acid substitutions. Amino acid substitutions selected using phylogenetic analysis were synthetically combined using a DOE design to create an information-rich set of gene variants, termed infologs. We used machine learning to identify and quantify protein sequence-function relationships against 14 different substrates. The resulting models were quantitative and predictive, serving as a guide for engineering of glutathione transferase activity toward a diverse set of herbicides. Predictive quantitative models like those presented here have broad applicability for bioengineering.
Comments: Infologs, Electra Cloning, ProteinGPS Protein Engineering