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A Novel Cytoplasmic Isopentenyl Diphosphate Isomerase Gene from Tomato (Solanum lycopersicum): Cloning, Expression, and Color Complementation

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Abstract

Isopentenyl diphosphate isomerase (IPI; EC5.3.3.2) catalyzes isomerization between isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), both of which are essential precursors for terpenoid biosynthesis. A novel gene encoding isopentenyl diphosphate isomerase (designated as SlIPI) was isolated from tomato based on tentative consensus (TC183769) and unigene SGN-U569721 sequences. The SlIPI cDNA contained a 708-bp open reading frame (ORF) encoding a 235-amino-acid protein. The deduced SlIPI protein had an isoelectric point of 5.06 and molecular weight of about 27.18 kDa. Amino acid sequence comparison analysis showed 83–95% similarity to IPIs from other plant species. Phylogenetic analysis revealed that SlIPI had the closest relationship to IPI from Nicotiana tabacum. The SlIPI was likely to be localized in cytoplasm; while, SlIPI2 contained a chloroplast transit peptide. A three dimensional structure modeling revealed that the structure of SlIPI was similar to that of SlIPI2. Tissue expression analysis indicated that SlIPI was constitutively expressed, with the highest expression level detected in the root. Heterologous expression of the recombinant SlIPI in engineered Escherichia coli resulted in the production and accumulation of carotenoid in E. coli, thus confirming that the SlIPI was a functional gene.

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Acknowledgments

This work was supported by grants from the Major State Basic Research Development Program (2009CB119005), the National Nature Science Foundation of China (No.30800756, No.30921002), and Xinjiang Corps Doctor Fund of China (No.2008JC04). We thank Dr. Jiancheng Zhang (Huazhong Agricultural University) for providing engineered plasmid pACCAR16∆crtX and Miss Xi Liu (Huazhong Agricultural University) for technical support.

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Authors and Affiliations

  1. National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, Hubei, 430070, China

    Jin Sun, Yu-Yang Zhang, Hui Liu, Zhe Zou, Chan-Juan Zhang, Xiao-Hui Zhang & Zhi-Biao Ye

  2. Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan, Hubei, 430070, China

    Hui Liu & Han-Xia Li

Authors
  1. Jin Sun
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  2. Yu-Yang Zhang
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  3. Hui Liu
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  4. Zhe Zou
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  5. Chan-Juan Zhang
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  6. Xiao-Hui Zhang
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  7. Han-Xia Li
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  8. Zhi-Biao Ye
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Corresponding author

Correspondence to Zhi-Biao Ye.

Additional information

Jin Sun and Yu-Yang Zhang contributed equally to this work.

Sequences data of SlIPI and SlIPI2 from this article have been deposited at GenBank under accession number EU253957 and GQ169536.

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Sun, J., Zhang, YY., Liu, H. et al. A Novel Cytoplasmic Isopentenyl Diphosphate Isomerase Gene from Tomato (Solanum lycopersicum): Cloning, Expression, and Color Complementation. Plant Mol Biol Rep 28, 473–480 (2010). https://doi.org/10.1007/s11105-009-0174-4

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