Figure 1

Construction of vectors and recombinant protein expression and purification. A. Schematic representation of recombinant pGEX4T1 vectors. Mage-a1 (aa118-219) gene segment, Hsp70 and Mage-a1-Hsp70 fusion gene were cloned into pGEX4T1 vector downstream of the GST sequence. B. GST protein was expressed by the vector pGEX4T-1 in E. coli Rosetta 2. M, protein molecular weight markers. Lanes 1–2, whole cell lysate without and with IPTG induction. Lane 3, purified GST protein. C. Mage-a1 protein expressed by recombinant vector pGEX4T-1-Mage-a1. M, protein molecular weight markers. Lanes 1–2, whole cell lysate without and with IPTG induction. Lanes 3–4, purified Mage-a1 protein at 2× or 1× loading concentration. D. Hsp70 and Mage-a1-Hsp70 protein expressed by recombinant vector pGEX4T-1-Hsp70 and pGEX4T-1-Mage-a1-Hsp70. M, protein molecular weight markers. Lanes 1–2, whole cell lysate of E. coli Rosetta 2 transformed with pGEX4T-1-Hsp70 without and with IPTG induction. Lanes 3–4, purified Hsp70 protein at 1× or 2× loading concentration. Lanes 5–6, whole cell lysate of E. coli Rosetta 2 transformed with pGEX4T-1-Mage-a1-Hsp70 without and with IPTG induction. Lanes 7–8, purified Mage-a1-Hsp70 protein at 1× or 2× loading concentration.