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Fig. 2 | Microbial Cell Factories

Fig. 2

From: Characterization and optimization of 5´ untranslated region containing poly-adenine tracts in Kluyveromyces marxianus using machine-learning model

Fig. 2

Evaluation of 5´ UTR poly(A)s through a dual-reporter system. (A) Flow chart of the dual-reporter system used to evaluate the effect of 5´ UTRs on protein production. Natural 5´ UTRs from abundant genes were randomly selected and cloned into separate vectors. The constructed plasmids were then transformed into K. marxianus, and the hosts were cultured separately for 72 h before FACS analysis. (B) Relative GFP abundance caused by 5´ UTRs. The relative GFP abundance was calculated as GFP/mCherry, where the relative GFP abundance caused by HXT4 5´ UTR was designated as 1. The mean relative GFP abundance of each 5´ UTR was ranked in ascending order, with the standard deviations (SD) shown (n = 3). The insert displayed 5´ UTRs that caused the lowest and highest relative GFP abundance. (C) Comparison of the relative GFP abundance caused by 5´ UTRs with poly(A) located at different positions. The significance was assessed using a two-tailed t-test. ****p < 0.0001; ns, p > 0.05

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Microbial Cell Factories

ISSN: 1475-2859

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