Molecular Breeding

, Volume 5, Issue 5, pp 453–461

Subcellular targeting of green fluorescent protein to plastids in transgenic rice plants provides a high-level expression system

  • In-Cheol Jang
  • Baek Hie Nahm
  • Ju-Kon Kim
Article

DOI: 10.1023/A:1009665314850

Cite this article as:
Jang, IC., Nahm, B.H. & Kim, JK. Molecular Breeding (1999) 5: 453. doi:10.1023/A:1009665314850

Abstract

In order to develop a high-level expression system in transgenic rice, we inserted a synthetic gene (sgfp) encoding a modified form of the green fluorescent protein (GFP) into two expression vectors, Act1-sgfp for an untargeted and rbcS-Tp-sgfp for a chloroplast targeted expression. Several fertile transgenic rice plants were produced by the Agrobacterium-mediated method. Confocal microscopic analyses demonstrated that, in cells expressing the Act1-sgfp, GFP fluorescence was localized within the cytoplasm and nucleoplasm whereas, in cells expressing the rbcS-Tp-sgfp fusion gene, the fluorescence was specifically targeted to chloroplasts and non-green plastids. The levels of sgfp expression were about 0.5% of the total soluble protein in mature leaf tissues of the Act1-sgfp transformed lines. In contrast, expression levels were markedly increased in mature leaf tissues of the rbcS-Tp-sgfp transformed lines, yielding about 10% of the total soluble protein. N-terminal sequencing of the localized GFPs revealed that the Tp-GFP fusion protein was correctly processed during import to non-green plastids, as well as to chloroplasts. Thus, our results demonstrate that GFP can be produced at high levels and localized in specific subcellular spaces of transgenic plants, providing a high-level expression system for general use in rice, an agronomically important cereal.

transgenic ricetransit peptideplastid targetinggreen fluorescent protein

Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • In-Cheol Jang
    • 1
  • Baek Hie Nahm
    • 1
  • Ju-Kon Kim
    • 1
  1. 1.Institute of Bioscience and BiotechnologyMyongji UniversityYongin, Kyonggi-DoKorea