Journal Article

Production of Polyhydroxybutyrate by Polycistronic Expression of Bacterial Genes in Tobacco Plastid

Yuko Arai, Toshiharu Shikanai, Yoshiharu Doi, Shigeo Yoshida, Isamu Yamaguchi and Hideo Nakashita

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 45, issue 9, pages 1176-1184
Published in print September 2004 | ISSN: 0032-0781
Published online September 2004 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pch139
Production of Polyhydroxybutyrate by Polycistronic Expression of Bacterial Genes in Tobacco Plastid

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  • Molecular and Cell Biology
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Transgenic techniques are used to enhance and improve crop production, and their application to the production of chemical resources in plants has been under investigation. To achieve this latter goal, multiple-gene transformation is required to improve or change plant metabolic pathways; when accomplished by plant nuclear transformation, however, this procedure is costly and time consuming. We succeeded in the metabolic engineering of the tobacco plant by introducing multiple genes within a bacteria-like operon into a plastid genome. A tobacco plastid was transformed with a polycistron consisting of the spectinomycin resistance gene and three bacterial genes for the biosynthesis of the biodegradable polyester, poly[(R)-3-hydroxybutyrate] (PHB), after modification of their ribosome binding sites. DNA and RNA analysis confirmed the insertion of the introduced genes into the plastid genome and their polycistronic expression. As the result, the transplastomic tobacco accumulated PHB in its leaves. The introduced genes and the PHB productivity were maternally inherited, avoiding genetic spread by pollen diffusion, and were maintained stably in the seed progeny. Despite the low PHB productivity, this report demonstrates the feasibility of transplastomic technology for metabolic engineering. This “phyto-fermentation” system can be applied to plant production of various chemical commodities and pharmaceuticals.

Keywords: Keywords: Biodegradable polyester — Metabolic engineering — Plastid transformation — Polycistron — Polyhydroxybutyrate.; Abbreviations: GC-MS, Gas chromatography-mass spectrometory; PCR, Polymerase chain reaction; PHB, poly-3-hydroxybutyrate; RBS, ribosome binding site.

Journal Article.  5981 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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