Applying the New Cloning Strategy: Production of a Specifically Designed PAA Diblock Containing a Zwitterionic Tail Block

6.1 Overview In Chapter 5 a new universal cloning strategy for the production of specifically designed surface-active PAA’s was introduced. Here, we apply this strategy to the production of a unique surface-active PAA designed to form brush layers on aluminum oxide surfaces. This chapter discusses the procedure taken in assembling the gene encoding for a PAA diblock consisting of a long zwitterionic tail block and a short acidic anchor block. Once the gene was constructed, the PAA was expressed with a HP-Thioredoxin (Trx) fusion tag in recombinant E. coli (see Figure 6.1). After purification, the Trx was removed and PAA product was isolated and its composition was confirmed. At the conclusion of this work, we will have demonstrated the application of the new cloning strategy towards the production of a truly unique PAA with a specified monomer sequence and composition. A manuscript submitted to Biomacromolecules based on the work presented in this chapter is located in Appendix A.