Figure 5.

Expression of the recombinant His-tagged Nt-inh1-encoded protein in E. coli and purification by Ni-affinity chromatography. The open reading frame of the Nt-inh1 cDNA was amplified by PCR and cloned into the pQE-30 vector (Qiagen). Expression of the fusion protein was induced with IPTG. A, Coomassie-stained SDS-PAGE gel. Lane M, Marker proteins; lane 1, protein from uninduced bacteria; lane 2, protein from IPTG-induced bacteria; lane 3, purified recombinant protein after Ni-affinity chromatography. B, Immunoblot of fractions 1 to 3 from A developed with the polyclonal antiserum directed against the INH protein (Krausgrill et al., 1996).