Translation controls the expression level of a chimaeric reporter gene
article
Transcriptional and translational fusions between the reading frame of the β-D-glucuronidase gene (gusA) and the 2′ as well as the 1′ promoter of mannopine synthase (mas), a TR locus of Agrobacterium tumefaciens, were made. The expression of these constructs was studied in the transgenic F1 offspring of independent tobacco transformants at the protein level by assaying for GUS activity and western blot analysis of the GUS protein and at the steady-state mRNA level. In leaves, stems and roots no correlation was found between steady-state levels of GUS mRNA and enzyme activity. In older tissues significantly higher GUS activities were found. This is explained by the stable character of the GUS protein together with an accumulation of protein upon ageing. Three to ten times higher GUS activities were found for in vitro grown plants than for greenhouse-grown plants of the same offspring, despite similar levels of GUS mRNA. Roots from in vitro grown plants display three to ten times higher GUS activities than stems and leaves. In transgenic plants grown in vitro, containing a translational fusion with two AUGs in phase, the initiation of translation in leaf material occurred at both AUGs. Initiation of translation at the first AUG, however, was ten times more frequent. In contrast, initiation in roots from in vitro grown plants occurred exclusively at the second AUG. © 1992 Kluwer Academic Publishers. Chemicals/CAS: beta glucuronidase, 9001-45-0; DNA, 9007-49-2; hydrolyase, 9044-86-4; Chimeric Proteins; DNA, 9007-49-2; Glucuronidase, EC 3.2.1.31; Hydro-Lyases, EC 4.2.1.; mannopine synthase, EC 4.2.1.-; Oligodeoxyribonucleotides; RNA, Messenger
Topics
β-D-glucuronidaseGene expressionInitiation of translationMannopine synthase promoterBeta glucuronidaseChimeric proteinDNAMannopine synthaseOligodeoxyribonucleotideAgrobacterium tumefaciensBiosynthesisEnzymologyGene expression regulationGenetic transcriptionGeneticsIsolation and purificationMetabolismMolecular geneticsNucleotide sequencePlantPromoter regionEstriction mappingRNA translationTobaccoTransgenic plantWestern blottingAgrobacterium tumefaciensBase SequenceBlotting, SouthernBlotting, WesternChimeric ProteinsDNAGene Expression Regulation, EnzymologicGlucuronidaseHydro-LyasesMolecular Sequence DataOligodeoxyribonucleotidesPlants, Genetically ModifiedPlants, ToxicPromoter Regions (Genetics)Reading FramesRestriction MappingRNA, MessengerSupport, Non-U.S. Gov'tTobaccoTranscription, GeneticTranslation, Genetic
TNO Identifier
231978
ISSN
01674412
Source
Plant Molecular Biology, 20(5), pp. 921-938.
Pages
921-938
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