In comparing Tsynth8 and Tsynth30 (Tmini) for use in yeast, both demonstrated increased heterologous protein expression and transcript levels greater than 2-fold over the commonly used TEF1 terminator [Curran et al.]. However, specifically for use in Y. lipolytica, Tsynth8 showed 3-fold greater protein expression than Tsynth30. In addition, Tsynth8 had slightly lower cryptic promoter activity and a lower tendency for transcript read-through, which would signal insufficient transcript termination. Most significantly, Tsynth8 could be ordered as a sequence from IDT; whereas Tsynth30, with it’s extended TA efficiency region, could not.

To confirm that the terminator Tsynth8 did have more protein expression in Y. lipolytica, we tried to replicate the experiment stated above. Where Tsynth8 should have 3-fold protein expression in Y. lipolytica. What we did is we inoculated Y. lipolytica with the terminator plasmids that had the gene hrGFP regulated by the following terminators: Tsynth8, Tsynth30, TEF1. These plasmids are the same exact plasmids that were used in the paper: Short Synthetic Terminators for Improved Heterologous Gene Expression in Yeast . Once inoculated, the cultures were diluted to the same OD600, and then every 2 hours their OD600 and fluorescence levels were taken. Our data for time versus OD600 can be seen in Fig.1, and the time vs fluorescence can be seen in Fig.2