Fully automated molecular biology: Plasmid-Based Functional Proteomic Workcell Evaluation and Characterization of Yeast Strains with Optimized "Trojan Horse" Amino Acid Scanning Mutational Inserts.

Authors: Hughes, Stephen; Qureshi, Nasib; Mertens, Jeffrey; Cotta, Michael; Hector, Ronald - Ron; Bischoff, Kenneth; Liu, Siqing; 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., Qureshi, N., Mertens, J.A., Cotta, M.A., Hector, R.E., Bischoff, K.M., Liu, S., Li, X-L., 2007. Fully automated molecular biology: Plasmid-based functional proteomic workcell evaluation and characterization of yeast strains with optimized "Trojan Horse" amino acid scanning mutational inserts [abstract]. PEPTALK 2007. Cambridge Healthtech Institute. Poster 509, p. 4.

Interpretive Summary:

Technical Abstract: The optimization of various genes is important in cellulosic fuel ethanol production from S. cerevisiae to meet the rapidly expanding need for ethanol derived from hemicellulosic materials. The United States Department of Agriculture has developed a fully automated platform for molecular biology routines that accomplishes screening of proteins in 96-well format via in vivo expression for evaluating growth on xylose using xylose isomerase expressing insecticidal peptides for use in crop killing insects, such as corn ear worms. These yeast will be used in cellulosic ethanol production and analyzed for optimum growth on pentose sugars such as xylose. 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 amino acid mutagenesis strategy using the ORF scanning method used at USDA. These operations allow rapid production of mutagenized insecticidal ORF inserts in pENTR D TOPO.