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Figure 3. Synthesis of GST-fusion mutant and wild-type amelogenins in vitro. (A) SDS-PAGE analysis of the recombinant GST-fusion products in the E. coli lysates. (B) Immunoblotting with anti-amelogenin antibody. Lane M: molecular markers. Other lanes indicate cell lysates of an E. coli strain containing the vacant expression vector (lanes 1,2), GST-tagged wild amelogenin (lanes 3,4), and GST-tagged mutant amelogenin (lanes 5,6). Blotting under conditions with (lanes 2,4,6) or without (lanes 1,3,5) isopropyl-ß-d-thio-galactopyranoside (ITPG)-induction. The faster-migrating bands in lanes 4 and 6 are suspected to be the cleavage products of GST-fusion amelogenin (wild or mutant) protein.
