Expression of a bacterial lysine decarboxylase gene and transport of the protein into chloroplasts of transgenic tobacco |
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Authors: | S Herminghaus P H Schreier J E G McCarthy J Landsmann J Botterman J Berlin |
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Institution: | (1) BBA-Institut für Biochemie, Biologische Bundesanstalt für Land- und Forstwirtschaft, Messeweg 11/12, D-3300 Braunschweig, Germany;(2) Institut für Biotechnologie PF-E, Bayer-AG, D-5090 Leverkusen, Germany;(3) GBF-Gesellschaft für Biotechnologische Forschung m.b.H., D-3300 Braunschweig, Germany;(4) Plant Genetic Systems NV, Gent, Belgium |
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Abstract: | A possible approach for altering alkaloid biosynthesis in plants is the expression of genes encoding key enzymes of a pathway such as lysine decarboxylase (ldc) in transgenic plants. Two strategies were followed here: one focused on expression of the gene in the cytoplasm, the other on subsequent targeting of the protein to the chloroplasts. Theldcgene fromHafnia alvei was therefore (a) placed under the control of the 1 promoter of the bidirectional Tr promoter fromAgrobacterium tumefaciens Ti- plasmid, and (b) cloned behind therbcS promoter from potato fused to the coding region of therbcS transit peptide. Bothldc constructs, introduced intoNicotiana tabacum with the aid ofA. tumefaciens, were integrated into the plant genome and transcribed as shown by Southern and northern hybridization. However, LDC activity was only detectable in plants expressing mRNA under the control of therbcS promoter directing the LDC fusion protein into chloroplasts with the aid of the transit peptide domain. In plants expressing the processed bacterial enzyme cadaverine levels increased from nearly zero to 0.3–1% of dry mass. |
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Keywords: | Nicotiana tabacum plant transformation gene expression bacterial lysine decarboxylase protein transport chloroplasts cadaverine |
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