Foot-and-mouth disease virus (FMDV) 2A constructs have been successfully used for the production of “Golden Rice”, a β-carotene producing rice strain. However, to allay public fears and opposition to plants carrying a mammalian pathogenic viral sequence, 2A-like synthetic sequences from Thosea asigna virus and Infectious myonecrosis virus were used to coordinate the coexpression of carotenoid biosynthetic genes. Here, up to four carotenogenic genes encoding PSY, CRTI, BCH and BKT were concatenated and produced β-carotene, zeaxanthin, and ketocarotenoids (astaxanthin and adonixanthin) in transgenic rice seeds displaying color variation due to the difference in carotenoid content and composition.
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Protein levels were first examined to compare the cleavage efficiency among T2A, I2A, and F2A using all bicistronic lines of stPTAC and stPIAC with a representative line of previous stPAC including a F2A (Fig. 2c, top). In an immune-blot with seed proteins (40 μg), T2A exhibited a 1.2- and 2.4-fold higher “cleavage” efficiency compared with I2A2 and F2A respectively. Moreover, T2A and I2A2 did not generate the any un-cleaved product, unlike F2A, which produced PSY:F2A:TP-CRTI – this suggests improved “cleavage” activity with T2A and I2A2. Further, PSY:2A forms between the stPTAC and stPIAC lines were similar using 2A antibody (Fig. 2c, bottom; Fig. 2d, left). It supported the better function of T2A than I2A2 contributed to the translation of the second CRTI ORF (Fig. 2c, top).This work was supported by grants from the Next-Generation BioGreen 21 Program (PJ01368801) and the Next Generation New Plant Breed Technology Program (PJ01477202) funded by the Rural Development Administration.
This work was supported by grants from the Next-Generation BioGreen 21 Program ( PJ01368801 ) and the Next Generation New Plant Breed Technology Program ( PJ01477202 ) funded by the Rural Development Administration .
© 2021 The Authors
- 2A peptide
- Metabolic engineering
ASJC Scopus subject areas
- Endocrinology, Diabetes and Metabolism
- Biomedical Engineering