Figure 3

Translational and transcriptional analysis of the Acthi gene in the HY and WT strains. (A) Western blotting analysis of ActhiS in the WT and HY strains. The protein samples were extracted on fermentation days 4 and 7. Fifteen micrograms of total protein was loaded for Western blotting analysis with anti-ActhiS antibodies. Actin was the loading control. ActhiS expression was higher in the HY strain compared to the WT strain. (B) The relative expression of Acthi in WT and HY strains. The RNA samples were extracted on fermentation days 4 and 7 and were subjected to RT-qPCR with Acthi and actin-specific primers, as described in the Methods. The relative mRNA abundance levels were calculated against actin gene levels. The histogram amplitude represents the average of three experiments. Error bars represent the standard deviations. A significant difference in the relative expression level of the Acthi gene in HY compared with that in WT is indicated as “*”; p value confidence levels: “**” < 0.01 and “***” < 0.001. Acthi was over-expressed in the HY stain throughout the fermentation process.