Genomics programs are discovering a large number of novel genes thus creating a need to express new proteins for therapeutics, diagnostics and to understand protein structure and function. LifeSensors proposes the creation of a new ubiquitin-fusion system to address this need. This proposal makes use of newly discovered regulatory properties of ubiquitin to secrete recombinant proteins into the medium. This fusion system will be tested in the baculovirus and yeast protein expression and secretion systems using known proteins of varied structure and function. It is expected that the improvements to the ub-fusion technology combined with secretion will significantly and reliably enhance the production and purification of biologically active proteins that have been difficult to express and purify in other systems. These results will be used in Phase II to develop a generic kit for expressing a variety of proteins or as part of specialized cDNA library expression systems.
Expression of biologically active proteins for research and biopharmaceutical purposes is hampered by the lack of a simple method that works for structurally diverse proteins in either eukaryotic or prokaryotic systems. We propose the use of ubiquitin-fusion technology combined with secretion to enhance the quantitative and qualitative expression and subsequent purification of biologically active proteins. The production of a kit incorporating these improvements could lead to a more reliable method of expressing and purifying proteins from all organisms.
| Marblestone, Jeffrey G; Edavettal, Suzanne C; Lim, Yiting et al. (2006) Comparison of SUMO fusion technology with traditional gene fusion systems: enhanced expression and solubility with SUMO. Protein Sci 15:182-9 |
| Butt, Tauseef R; Edavettal, Suzanne C; Hall, John P et al. (2005) SUMO fusion technology for difficult-to-express proteins. Protein Expr Purif 43:1-9 |
