Figure 4
Expression of the synthetic and wt recombinant taqIIRM gene variants. (A) Expression of the wt- recombinant taqIIRM gene. Lanes M, protein marker (Thermo Fisher Scientific/Fermentas); lane 1, control culture – crude lysate from E. coli expressing the cloned wt-taqIIRM gene, without induction (OD600 = 0.95); lane 2, control culture after 19 h of cultivation; lane 3, crude lysate from E. coli expressing the cloned wt-taqIIRM gene, before induction (OD600 = 0.9); lane 4, 1 h after induction; lane 5, 2 h; lane 6, 3 h; lane 7, 19 h; lane 8, purified, homogeneous recombinant syn-TaqII protein. (B) Expression of the recombinant syn-taqIIRM gene. Lanes M, protein marker (Thermo Fisher Scientific/Fermentas); lane 1, control culture – crude lysate from E. coli expressing the cloned syn-taqIIRM gene, without induction (OD600 = 0.95); lane 2, control culture after 19 h of cultivation; lane 3, crude lysate from E. coli expressing the cloned syn-taqIIRM gene, before induction (OD600 = 0.95); lane 4, 1 h after induction; lane 5, 2 h; lane 6, 3 h; lane 7, 19 h; lane 8, purified, homogeneous recombinant syn-TaqII protein. (C) Expression of the native taqIIRM gene in T. aquatiqus. Lane M, protein marker (GE Healthcare, Little Chalfont, United Kingdom); lane 1, purified, homogeneous recombinant syn-TaqII protein; lane 2, crude lysate from T. aquatiqus. (D) The amount of TaqII protein produced by E. coli cells expressing taqIIRM gene variants versus protein yield from native source.