FIG. 2.

Production and confirmation of antigenicity of TCoV N recombinant fusion protein produced by E. coli BL-21 transfected with pGEX-4T3-TCoV-N using 12% discontinuous SDS-PAGE (lanes 1 to 3) and subsequent Western blotting with anti-GST and anti-TCoV N-protein MAbs 1.01 and 4.23 (lanes 4 to 7). Lane M, molecular mass marker in kilodaltons; lane 1, nonpurified bacterial lysate after induction with IPTG; lane 2, TCoV N fusion protein purified with GST beads from bacterial lysate after induction with IPTG; lane 3, the same as lane 2 but from noninduced bacterial culture; lanes 4 and 5, Western blot using anti-GST antibodies against the TCoV N fusion protein expressed and purified as in lanes 2 and 3, respectively; lanes 6 and 7, Western blot using MAbs 1.01 and 4.23, respectively, against the TCoV N fusion protein expressed and purified as described for lane 2.