A Novel Technology Platform for Protein Engineering and Expression

University Of Texas At Austin, Austin TX

Investigators

Abstract

Many biotechnology applications related to secreted proteins have relied almost exclusively on the sec pathway mode of protein transport. However, many recombinant proteins are incompatible with secretion via the sec pathway for various reasons. The Principal Investigator (PI) proposes research to overcome the limitations of secretory protein expression in bacteria by capitalizing on the remarkable properties of the Tat secretion pathway. The PI proposes to exploit the unique features of the Tat pathway to overcome the limitations associated with secretory protein production and to develop completely new technology platforms for combinatorial protein screening and analysis. It is proposed to: (1) develop a useful technology for the high level expression of secretory proteins via the Tat pathway, specifically the PI will: (i) employ genetic means to identify leader peptides that maximize the flux of proteins exported though the Tat pathway; (ii) utilize cell engineering methods to further optimize protein secretion and (iii) examine in detail the effect of growth conditions on secretory protein production, (2) develop methods for the display of protein libraries by utilizing the Tat pathway and then investigate the hypothesis that altering the pathway of protein secretion will result in the isolation of additional and/or novel proteins capable of performing a desired function, and (3) develop a new technology for the detection of protein-protein interactions analogous to the yeast two-hybrid system by exploiting the Tat pathway.