4.4 Article

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

Journal

PLANT MOLECULAR BIOLOGY REPORTER
Volume 28, Issue 3, Pages 473-480

Publisher

SPRINGER
DOI: 10.1007/s11105-009-0174-4

Keywords

Isopentenyl diphosphate isomerase; Cloning; Functional characterization; Tomato (Solanum lycopersicum)

Funding

  1. Major State Basic Research Development Program [2009CB119005]
  2. National Nature Science Foundation of China [30800756, 30921002]
  3. Xinjiang Corps Doctor Fund of China [2008JC04]

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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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