Construction, Expression, and Characterization of Thymosin Alpha 1 Tandem Repeats in Escherichia coli

Figure 2

Expression, purification, and identification of Tα1③. (a) Expression of trx-Tα1③ in TOP10. Lane 1: protein marker; lane 2: total bacterial proteins of pThioHisA-Tα1③/TOP10 without induction; lane 3: total bacterial protein with IPTG induction. (b) Chromatogram of Q-Sepharose Fast Flow chromatography for purification of trx-Tα1③. The arrow indicated trx-Tα1③. (c) Chromatogram of SP-Sepharose Fast Flow chromatography for purification of Tα1③. The arrow indicated Tα1③. (d) SDS-PAGE analysis of trx-Tα1③ purification. Lane 1–3: total proteins of pThioHisA-Tα1③/TOP10 after IPTG induction (1 h, 3 h, 5 h); lane 4: supernatant of lysate heated at 80°C for 10 min; lane 5: purified trx-Tα1③. (e) Tricine-SDS-PAGE analysis of Tα1③ purification. Lane 1: standard peptide marker; lane 2: cleavage products without Tα1③; lane 3: purified Tα1③. (f) Western-blot analysis of trx-Tα1③. Lane 1: total bacterial proteins after IPTG induction; lane 2: purified trx-Tα1③.