The Development of a New Cloning Strategy for the Biosynthetic Production of Brush-Forming Poly(Amino Acids)
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We introduce here the development of a simple and versatile strategy for producing de novo-designed, high molecular weight PAA's using recombinant DNA technology. The basis of this strategy is that small DNA modules encoding for short PAA blocks can be easily inserted directly into a commercially available and unmodified expression vector. The insertions can be made repeatedly until the gene encodes for a polymer of desired molecular weight and composition. Thus, sequential modifications can be made to the PAA without having to re-start the gene assembly process from the beginning, thereby allowing for quick determination of how these changes affect polymer structure and function. The feasibility and simplicity of this method was shown during the production of a PAA, consisting of a long zwitterionic tail block and a short acidic anchor block, designed to form optimal brush layers on alumina surfaces. The success and flexibility of this method indicates that it can be applied for production of de novo-designed polypeptides in general. It is hoped that this method will contribute towards the rapid development of bio-inspired protein-based polymers for a variety of applications.
This dissertation also contains research that aimed to use phage display technology to develop a new liposome-based immunoassay against biological toxins. This work was part of a collaboration effort with the U.S. Department of Defense and Luna Innovations.
- Doctoral Dissertations