BioDesign Research / 2022 / Article / Fig 2

Research Article

Making Use of Plant uORFs to Control Transgene Translation in Response to Pathogen Attack

Figure 2

uORFs of ACD11 are cis-acting elements that repress downstream translation. (a) Diagram of three uORFs identified in the 5-UTR of ACD11. The main ORF (mORF) indicates ACD11. (b) Repression of GFP production by the 5-UTR of ACD11. Photos were taken 48 hours post agro-infiltration. Illustration of the indicated plasmids was shown below. (c) Relative fluorescence of leaves expressing indicated plasmids. Relative fluorescence was calculated based on the formula shown in Figure 1(c) (; , Student’s -test, ). (d) Relative transcript accumulation levels of GFP. Transcript accumulation levels of GFP were analyzed by qRT-PCR with NbELF18 as internal reference. Bars represent standard errors from three biological replicates (; ; , ; Student’s -test). (e) Decreased GFP protein expression confirmed by Western blot assay. The GFP protein accumulation was detected by α-GFP antibody. Equal loading of each sample is indicated by the luciferase protein. (f) ACD11 uORFs act in cis. uORFs was expressed in trans in leaves expressing uorf1/2/3 plasmid. Relative fluorescence was calculated based on the formula shown in Figure 1(c) (; , Student’s -test, ).