|Li, Xin Liang|
Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 1/12/2007
Publication Date: 1/12/2007
Citation: Hughes, S.R., Li, X., Liu, S., Bischoff, K.M., Mertens, J.A., Qureshi, N., Riedmuller, S., Bartolett, S., Farrelly, P., Panavas, T., Butt, T., Cotta, M.A. 2007. Fully automated molecular biology routines on a plasmid-based functional proteomic workcell: Evaluation and characterization of yeast strains optimized for growth on xylose and engineered to express an insecticidal peptide [abstract]. PEPTALK 2007, Cambridge Healthtech Institute. Talk 83, p. 7.
Technical Abstract: Optimization of genes important to production of fuel ethanol from hemicellulosic biomass for use in engineering improved commercial yeast strains is necessary to meet the United States' rapidly expanding need for ethanol. United States Department of Agriculture, Agricultural Research Service, National Center for Agricultural Utilization Research has developed a fully automated robotic platform for molecular biology routines that can accomplish rapid screening of proteins in 96-well format via an in vivo expression system to evaluate yeast strains expressing xylose isomerase and optimized for growth on xylose. 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 and hemicellulosic 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-ARS-NCAUR. These operations allow rapid and efficient production and screening of mutagenized insecticidal peptides for optimum expression in yeast strains using xylose as a feedstock.