dc.contributor.author	Henderson, Douglas Brian	en
dc.contributor.committeechair	Davis, Richey M.	en
dc.contributor.committeemember	Ducker, William A.	en
dc.contributor.committeemember	Goldstein, Aaron S.	en
dc.contributor.committeemember	Williams, Kimberly Forsten	en
dc.contributor.committeemember	Van Cott, Kevin E.	en
dc.contributor.department	Chemical Engineering	en
dc.date.accessioned	2014-03-14T20:20:20Z	en
dc.date.adate	2004-12-17	en
dc.date.available	2014-03-14T20:20:20Z	en
dc.date.issued	2004-12-10	en
dc.date.rdate	2007-12-17	en
dc.date.sdate	2004-12-15	en
dc.description.abstract	The design and discovery of new surface-active polymers that self-assemble on solid substrates to form brush layers will have a major impact on numerous applications. Through recombinant DNA technology, there exists the potential to harness a cell's protein synthesis machinery to produce a brush-forming poly(amino acid) (or PAA) with an exactly specified amino acid sequence, thus controlling the polymer's composition at a level unequaled by conventional organic polymer synthesis. The presented work demonstrates the cloning, expression, purification and characterization of de novo-designed PAA's designed to form brush layers on alumina surfaces. Using conventional recombinant DNA methods, the feasibility of producing a PAA consisting of a poly-glutamate block and a poly-proline block was demonstrated. However, the PAA design was limited by the inherent limitations of conventional cloning techniques. 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.	en
dc.description.degree	Ph. D.	en
dc.identifier.other	etd-12152004-145307	en
dc.identifier.sourceurl	http://scholar.lib.vt.edu/theses/available/etd-12152004-145307/	en
dc.identifier.uri	http://hdl.handle.net/10919/30103	en
dc.publisher	Virginia Tech	en
dc.relation.haspart	10Chapt_8_Phage_Display_Work.pdf	en
dc.relation.haspart	11Appendix_A_Cloning_Method_Manuscript.pdf	en
dc.relation.haspart	09Chapt_7_Conclusions_and_Future_Work.pdf	en
dc.relation.haspart	08Chapt_6_Application_of_New_Cloning_Strategy.pdf	en
dc.relation.haspart	12Vita.pdf	en
dc.relation.haspart	07Chapt_5_Development_of_New_Cloning_Strategy.pdf	en
dc.relation.haspart	02Abstr_TOC_Fig_Tab_Abbr.pdf	en
dc.relation.haspart	06Chapt_4_A_Brush-Forming_PAA.pdf	en
dc.relation.haspart	05Chapt_3_1st_Generation_PAA.pdf	en
dc.relation.haspart	04Chapt_2_Backgound.pdf	en
dc.relation.haspart	03Chapt_1_Project_Summary.pdf	en
dc.relation.haspart	01Title_page.pdf	en
dc.rights	In Copyright	en
dc.rights.uri	http://rightsstatements.org/vocab/InC/1.0/	en
dc.subject	brush	en
dc.subject	recombinant DNA	en
dc.subject	poly(amino acids)	en
dc.subject	self-assembly	en
dc.subject	biopolymer	en
dc.title	The Development of a New Cloning Strategy for the Biosynthetic Production of Brush-Forming Poly(Amino Acids)	en
dc.type	Dissertation	en
thesis.degree.discipline	Chemical Engineering	en
thesis.degree.grantor	Virginia Polytechnic Institute and State University	en
thesis.degree.level	doctoral	en
thesis.degree.name	Ph. D.	en

The Development of a New Cloning Strategy for the Biosynthetic Production of Brush-Forming Poly(Amino Acids)

Files

Original bundle

Collections