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Expression of Recombinant tryptophan decarboxylase in Different Subcellular Compartments in Tobacco Plant

Acta Botanica Sinica 2002年第3期44卷 314-317页

作者：王淼李秋荣 Stefano Di FIORE Rainer FISCHER 中国科学院沈阳应用生态研究所沈阳110016 沈阳军区总医院沈阳110016 Botanical Institute of Rheinisch-Westflisch Technische Hochschule Aachen

The gene encoded for tryptophan decarboxylase (TDC), which is the key enzyme in terpenoil indole alkaloids pathway, was targeted to different subcellular compartments and stably expressed in transgenic tobacco (Nicotiana tabacum L.) plants at the levels detected by Western blot and tryptamine accumulation analysis. It was shown that the TDC was located in subcellular compartments, the chloroplasts and cytosol. The recombinant TDC targeted to chloroplasts and cytosol in tobacco plants was effectively expressed as soluble protein by Western blot analysis and enzymatic assay. The level of tryptamine accumulation in chloroplast was higher than that in cytosol and very low in vacuole and endoplasmic reticulum (ER) to be hardly detected by Western blot analysis. It was indicated that the highest amount of tryptamine was in chloroplasts, lower in endoplasmic reticula and the lowest in vacuoles as compared to those in wild type plants. The TDC targeted to different subcellular compartments of tobacco plants and its expression level were studied by different nucleotide sequences coding signal peptides at 5'-end of tdc gene in order to know the effects of the TDC in compartmentation on its functionality.

关键词： tryptophan decarboxylase alkaloids biosynthetic pathway Nicotiana tabacum subcellular compartment

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