| mRNA display is a powerful in vitro protein evolution technique to evolve functional proteins and peptides. The covalent bond between the genotype, phenotype and cell-free process enable it to work on libraries with a larger size than other display technologies. Besides the successful application to functional protein selections, the controlled conditions of mRNA display also provide implications to the understanding of natural protein evolution. This work includes the structural and functional analysis of a non-biological ATP-binding protein derived from mRNA display, the selection of transferrin-binding peptides from a random library, the optimization of transferrin-binding peptides from a small library screen and the development of a semi-rational approach to novel protein interface design in cooperation with the computational protein design strategy. Functional protein and peptides were obtained in these projects. Structural and functional analysis of these proteins provides information to help understand the basic principles of natural protein evolution. |
