Expression analysis of Ala35Thr mutated FAH. Using the TnT Coupled Reticulocyte Lysate System (Promega) (2), wild-type FAH and mutated FAH were expressed. An autoradiography of 35S-methionine-labeled translation products separated by polyacrylamide gel electrophoresis is shown. Lanes 1 and 7: luciferase expression (control). Lanes 2 and 6: expression of 43 kDa FAH control. Lane 3: expression of FAH mutant with Ala35Thr. Lane 4: plasmid without insert. Lane 5: no DNA control. Ala35Thr mutation of the FAH gene leads to absence of protein expression.