II.A.h.1 - Biochemistry: Identification and evolution of adhesive peptides with novel specificity using yeast surface display
Abstract
The objective of the research project is to develop a yeast-based system for the generation and selection of peptide sequences with novel adhesive properties. Specifically, yeast display technology will be enhanced by introducing: 1) a novel mechanism for imparting genetic diversity and 2) a non-canonical amino acid incorporation technique to impart chemical modifications into putative adhesive proteins. The research effort will encompass three specific aims: 1) Measure the adhesive capability of peptides derived from marine animals using yeast display. A yeast display system will be developed to characterize and identify adhesive proteins. Known adhesive proteins will be cloned and expressed and their properties will be characterized. Molecular techniques will be used to identify putative adhesive proteins from the limpet Crepidula fornicata. This goal is significant in that it will establish the budding yeast S. cerevisiae as a system for the screening of adhesive peptides. 2) Design a system for generating genetic diversity within repetitive coding regions in yeast. A primary limitation of yeast display and other display methodologies is the dearth of mechanisms for continuous molecular diversification. Recent results have identified features within repetitive coding regions that confer higher rates of genetic instability. The DNA sequence characteristics that underlie this instability will be elucidated, and this diversity-generating mechanism will be applied in the yeast display system to evolve novel adhesive peptides derived from the repetitive adhesive domains of marine animals. 3) Incorporate DOPA into adhesive peptides in yeast. The yeast display system will be engineered to measure the influence of DOPA incorporation on adhesion and to test the limits of DOPA incorporation into proteins produced in yeast.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Jan 12, 2017
- Source ID
- W911NF1610175
Entities
People
- Stephen M. Fuchs
Organizations
- Army Contracting Command
- Tufts University
- United States Army