Secondary Structure Prediction of Protein Constructs Using Random Incremental Truncation and Vacuum-Ultraviolet CD Spectroscopy
Fig 2
Screening expression level and solubility of UDE truncation libraries.
(A) Size fractionation of UDE fragments generated by unidirectional truncation on agarose gel. In the lanes next to the DNA ladders is the vector with total length UDE gene at higher position while the empty vector is at a lower position. N1–N3 and C1–C3 marked samples show by the exonuclease III truncation generated UDE constructs. (B) Assessment of UDE sublibraries size and diversity by PCR screen. (C) Separation of purified protein fractions on Ni2+-NTA resin from N-terminal (upper panels) and C-terminal (bottom panels) libraries on SDS-PAGE.