Figure 9.

The effect of the 5′-UTRs on the translational efficiency is conserved in different plant species. (A) GFP levels in Arabidopsis protoplasts. Reporter constructs of the AT1G58420 or AT1G04850 5′-UTRs were co-transformed into Arabidopsis protoplasts together with a GUS construct and the expression levels of GFP and GUS were determined by western blot analysis using anti-GFP and anti-GUS antibodies, respectively. (B) Quantification of translational efficiency. To quantify the translational efficiency, the signal intensity of the immunoblots in (A) was quantified using the multi-gauge software equipped to the LAS3000 and the GFP levels were normalized using the GUS levels. Error bar, standard deviation (n = 3). (C) GFP levels in tobacco leaf tissues. The GFP constructs with the 21 nt region of AT1G58420 or AT1G04850 5′-UTRs in a binary vector were transformed into Nicotiana benthamiana leaf tissues by Agrobacterium-mediated infiltration. The expression levels of GFP and HPT were determined by western blot analysis using anti-GFP and anti-HPT antibodies, respectively. HPT was used to normalize the transformation efficiency. (D) Quantification of translational efficiency. To quantify the translational efficiency, the signal intensity of the immunoblot in (C) was quantified using the multi-gauge software equipped to the LAS3000 and the GFP levels were normalized using the HPT levels. Error bar, standard deviation (n = 3).