Ubiquitin promoter–terminator cassette promotes genetically stable expression of the taste-modifying protein miraculin in transgenic lettuce

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Lettuce is a commercially important leafyvegetable that is cultivated worldwide, and it is also atarget crop for plant factories. In this study, lettuce wasselected as an alternative platform for recombinant miraculinproduction because of its fast growth, agronomicvalue, and wide availability. The taste-modifying proteinmiraculin is a glycoprotein extracted from the red berries ofthe West African native shrub Richadella dulcifica.Because of its limited natural availability, many attemptshave been made to produce this protein in suitable alternativehosts. We produced transgenic lettuce with miraculingene driven either by the ubiquitin promoter/terminator cassette from lettuce or a 35S promoter/nosterminator cassette. Miraculin gene expression and miraculinaccumulation in both cassettes were compared byquantitative real-time PCR analysis, Western blotting, andenzyme-linked immunosorbent assay. The expression levelof the miraculin gene and protein in transgenic lettuce washigher and more genetically stable in the ubiquitin promoter/terminator cassette than in the 35S promoter/nosterminator cassette. These results demonstrated that theubiquitin promoter/terminator cassette is an efficient platformfor the genetically stable expression of the miraculinprotein in lettuce and hence this platform is of benefit forrecombinant miraculin production on a commercial scale.