Generation of Polyprotein Constructs

The talin fragment polyprotein constructs, including flanking I₂₇, were synthesized and cloned in to pFN18a. The polyproteins were expressed in E. coli BL21-CodonPlus (DE3)-RILP competent cells, using the T7 promoter present in the plasmid. Protein expression was induced with IPTG when the culture reached an OD_600 nm of 0.6. Cells were lysed with lysozyme and sonication before being purified with Ni-NTA beads in a batch process. The eluted proteins were analyzed for purity with SDS-PAGE. Final concentration of protein used for experiments was 1–10 μg/mL.