ARS | Publication request: Fully automated molecular biology routines on a plasmid-based functional proteomic workcell: Evaluation and Characterization of Yeast Strains Optimized for Growth on Xylose Expressing "Stealth" Insecticidal Peptides.

Submitted to: Meeting Abstract
Publication Type: Abstract
Publication Acceptance Date: March 29, 2007
Publication Date: March 29, 2007
Citation: Hughes, S.R., Li, X., Hector, R.E., Bischoff, K.M., Cotta, M.A., Mertens, J.A., Dowd, P.F., Johnson, E.T. 2007. Fully automated molecular biology routines on a plasmid-based functional proteomic workcell: Evaluation and Characterization of Yeast Strains Optimized for Growth on Xylose Expressing "Stealth" Insecticidal Peptides [abstract]. ACS National Meeting. p. 108.

Technical Abstract: Optimization of genes important to production of fuel ethanol from hemicellulosic biomass for use in developing improved commercial yeast strains is necessary to meet the rapidly expanding need for ethanol. The United States Department of Agriculture has developed a fully automated platform for molecular biology routines that accomplishes rapid screening of proteins in 96-well format via in vivo expression to evaluate yeast strains optimized for growth on xylose by addition of xylose isomerase. The yeast strains are also engineered to simultaneously express insecticidal peptides toxic to crop pests, such as corn ear worms. These yeast strains will be used in cellulosic ethanol production and at the same time provide a vehicle to deliver pesticide to crops for ingestion by insect pests. Here we discuss all operations of the automated workcell, including the various expression protocols for profiling and characterization of insecticidal peptides and optimized open reading frames generated by an amino acid scanning mutagenesis strategy used at USDA. These operations allow rapid and efficient production and screening of mutagenized insecticidal peptides for optimum expression in yeast strains using xylose as a feedstock.