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Evaluation of seed storage-protein gene 5′ untranslated regions in enhancing gene expression in transgenic rice seed

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Abstract

5′ untranslated regions (UTRs) are important sequence elements that modulate the expression of genes. Using the β-glucuronidase (GUS) reporter gene driven by the GluC promoter for the rice-seed storage-protein glutelin, we evaluated the potential of the 5′-UTRs of six seed storage-protein genes in enhancing the expression levels of the foreign gene in stable transgenic rice lines. All of the 5′-UTRs significantly enhanced the expression level of the GluC promoter without altering its expression pattern. The 5′-UTRs of Glb-1 and GluA-1 increased the expression of GUS by about 3.36- and 3.11-fold, respectively. The two 5′-UTRs downstream of the Glb-1, OsAct2 and CMV35S promoters also increased GUS expression level in stable transgenic rice lines or in transient expression protoplasts. Therefore, the enhancements were independent of the promoter sequence. Real-time quantitative RT-PCR analysis showed that the increase in protein production was not accompanied by alteration in mRNA levels, which suggests that the enhancements were due to increasing the translational efficiencies of the mRNA. The 5′-UTRs of Glb-1 and GluA-1, when combined with strong promoters, might be ideal candidates for high production of recombinant proteins in rice seeds.

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Acknowledgments

This work was supported by the National Program of Transgenic Variety Development of China (2008ZX08009-004), the Ministry of Science and Technology of China (No. 2007DFA3077) and the Natural Science Foundation of China (No. 30871375). The Glb-1 promoter was kindly provided by Dr. F. Takaiwa, NIAS, Japan.

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  1. Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing, 100093, China

    Wen Xian Liu, Hua Liang Liu, Zhi Jian Chai, Xiu Ping Xu, Yan Ru Song & Le Qing Qu

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  1. Wen Xian Liu
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  2. Hua Liang Liu
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  3. Zhi Jian Chai
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  4. Xiu Ping Xu
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  5. Yan Ru Song
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  6. Le Qing Qu
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Correspondence to Le Qing Qu.

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Communicated by E. Guiderdoni.

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Liu, W.X., Liu, H.L., Chai, Z.J. et al. Evaluation of seed storage-protein gene 5′ untranslated regions in enhancing gene expression in transgenic rice seed. Theor Appl Genet 121, 1267–1274 (2010). https://doi.org/10.1007/s00122-010-1386-6

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  • DOI: https://doi.org/10.1007/s00122-010-1386-6

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