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Characterization of the rice RbcS3 promoter and its transit peptide for use in chloroplast-targeted expression

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Abstract

Here, we report characterization of an RbcS promoter and its transit peptide for use in chloroplast-targeted expression. RbcS3 that encodes a small subunit of ribulose bisphosphate carboxylase/oxygenase is of great interest due to its green tissue-specific expression such as leaf and leaf sheaths. We isolated the RbcS3 promoter and its transit peptide sequence and linked to GFP for characterization in transgenic plants. Our qRT-PCR analysis on leaves and roots of transgenic plants revealed that GFP transcript levels were higher by fivefold in leaves of RbcS3:TP3:GFP transgenic lines than in those of the RbcS3:GFP transgenic lines, suggesting that the transcript levels were increased by chloroplast-targeted expression, as shown in previously characterized RbcS1:TP1 system. The transit peptide sequence TP3 was sufficient to target GFP to chloroplasts as observed in protoplasts of RbcS3:TP3:GFP transgenic lines. GFP fluorescence in RbcS3:GFP plants was detected only in the cytosol. The GFP protein levels in leaves of RbcS3:TP3:GFP plants were 0.12–0.23 % of the total soluble leaf proteins. These levels were higher or comparable to those of PGD1:GFP plants (GFP driven by a constitutive promoter PGD1). Taken together, these results indicate that the fusion of the RbcS3 promoter to its transit peptide, RbcS3:TP3, provides a chloroplast-targeting expression system.

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Acknowledgments

This research was supported by the Rural Development Administration under the Next-Generation BioGreen 21 Program (Project No. PJ011135 to J.-K.K.). This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT and Future Planning (Project No. R1A2A1A11051690 to J.-K.K., Project No. R1A1A3012706 to S.-H.P. and Project No. R1A1A1012877 to J.S.J).

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    Authors and Affiliations

    1. Crop Biotechnology Institute, GreenBio Science and Technology, Seoul National University, Pyeongchang, 25354, Korea

      Su-Hyun Park, Jin Seo Jeong, Yang Do Choi & Ju-Kon Kim

    2. Department of Agricultural Biotechnology, Seoul National University, Seoul, 151-921, Korea

      Yang Do Choi

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    1. Su-Hyun Park
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    2. Jin Seo Jeong
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    3. Yang Do Choi
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    Correspondence to Ju-Kon Kim.

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    S.-H. Park and J. S. Jeong contributed equally to this work.

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    Park, SH., Jeong, J.S., Do Choi, Y. et al. Characterization of the rice RbcS3 promoter and its transit peptide for use in chloroplast-targeted expression. Plant Biotechnol Rep 9, 395–403 (2015). https://doi.org/10.1007/s11816-015-0376-1

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    • DOI: https://doi.org/10.1007/s11816-015-0376-1

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