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. 2013 Sep 30;42(1):485–498. doi: 10.1093/nar/gkt864

Figure 2.

Figure 2.

The 21 nt region immediately upstream from the AUG start codon of various Arabidopsis genes shows a wide variation in translational efficiency. (A) Schematic presentation of fusion constructs. 5′-UTR, the 5′-untranslated region (165 nts) from the expression vector; 21 nt, 21 nt-long 5′-UTRs from various Arabidopsis genes; GFP-coding region; 3′-UTR, 3′-untranslated region from the expression vector. (B) Translation efficiency of the 5′-UTRs of various Arabidopsis genes. Reporter constructs were introduced into protoplasts together with a reference construct GUS and total protein extracts were subjected to western blot analysis using anti-GFP and anti-GUS antibodies. The 5′-UTR of Rbcs1A was used as a reference for the expression. (C) Quantification of translational efficiency. To quantify the translational efficiency, the signal intensity of immunoblots in (B) was quantified using the multi-gauge software equipped to the LAS3000 (FUJIFILM) and the GFP levels were normalized with the GUS level. (D) Quantification of transcriptional efficiency. To quantify the transcriptional efficiency, total RNA from the protoplasts transformed with two representative 5′-UTR constructs (asterisk in (C)) was used for qRT-PCR using specific primers for the GFP-coding region. The GUS transcript level was used as reference.