Engineering and characterization of single chain antibody fragments (scFvs) specific to key enzymes in polyamine biosynthesis and manipulation of polyamine pathway by constitutive expression of recombinant ODC and SDE enzymes in transgenic tobacco

TL;DRAbstract

Polyamines are ubiquitous low-molecular weight polycationic compounds found in all living organisms and involved in crucial developmental, physiological and metabolic processes. The majority of information available about the polyamine pathway was obtained by using a wide range of metabolic inhibitors (Evans and Malmberg, 1989; Tiburcio et al., 1990) and modulation of specific enzymatic activities via heterologous gene expression in transgenic plants. Previous studies have indicated that the polyamine pathway is tightly controlled in the end product accumulation. The objective of this study was to achieve changes in the pathway by combining genetic engineering and antibody technology approaches. ODC a key enzyme in the polyamine biosynthesis, was amplified from a human prostate cDNA library and subcloned into plant expression vector for transient expression in the apoplast, cytosol and endoplasmatic reticulum. ODC was accumulated to higher levels in plant cytosol and apoplast, while OD

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