FIG. 7.
(A) The double mutation of mup1-100 and sam1-224 increased SeMet resistance of S. cerevisiae. A serial dilution of TKY371-4A (mup1-100 sam1-224) cells harboring various combinations of pRS315-MUP1, pRS316-SAM1, and empty vectors was spotted on SC-Met/Leu/Ura medium with or without 0.05 mM SeMet for 1 day at 30°C. (B) Tricine-SDS-PAGE analysis of the L123M peptide expressed in the presence of 0.25 mM SeMet by SeMet-resistant mutants. The samples from Ni-NTA affinity chromatography were separated on a Tricine gel (15 to 20%; Wako), and protein bands were visualized by silver staining. The OD600 values at induction and harvest are indicated.