Fig. 4

Relative plasmid gene copy-number determined from real-time PCRs. The transformed yeast were inoculated at OD₆₀₀ = 0.15 into BMMD SFC without or with 1000 µM methionine and grown for 24 h. The cells were harvested and total DNA was isolated from 10 mL cultures. The plasmid names refer to the gap–repair plasmids containing the expression cassettes with the promoter and the expressed protein indicated below, all of which are present in the final pSAC35–based expression plasmid in yeast. Cultures were grown without and with (1000 µM) methionine in the initial media. Error bars represent the standard deviation of triplicate analysis of triplicate experimental cultures (n = 9). The fold change is relative to DB1 [pDB4351] without methionine. FLP1 was used as the single-copy plasmid gene and TAF10 was used as the genomic control