Submitted to: Great Lakes Regional American Chemical Society Symposium
Publication Type: Abstract Only
Publication Acceptance Date: 10/20/2004
Publication Date: 10/20/2004
Citation: Hughes, S.R., Li, X., Cotta, M.A., Bischoff, K.M., Riedmuller, S., Farrelly, P. 2004. A plasmid-based functional proteomic workcell used to optimize open reading frames and generate improved strains for commercial use [abstract]. 36th Great Lakes Regional American Chemical Society Symposium, October 17-20, 2004, Peoria, Illinois. Poster 361. Interpretive Summary:
Technical Abstract: We are developing a high throughput plasmid-based functional proteomic workcell that integrates in a robotic platform the operations of media filling, colony picking, plate sealing, advanced liquid handling, incubation and shaking, and assay data collection into a custom database, with 2D bar code tracking of plates, clones, strains, and datapoints, for rapid optimization of clones and expression of full-length cDNA libraries to produce improved bacterial and yeast strains. In producing the plasmid-based libraries of full-length cDNAs that are the source of the optimized sequences for expression in yeast and bacteria, the robotic platform must be capable of running in vitro and in vivo expression reactions to obtain proteins in proper conformation so that clones having optimal traits can be identified before placement into these strains permanently. Using this high throughput platform, the yeast and bacteria strains can be screened for growth characteristics and production of byproducts and biocatalysts via standard biochemical assays, and this presentation will discuss the design, construction, and validation of these systems.