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OtherMOLECULAR BIOLOGY AND GENE REGULATION
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cDNA Cloning of Carrot (Daucus carota) Soluble Acid [beta]-Fructofuranosidases and Comparison with the Cell Wall Isoenzyme

C. Unger, M. Hardegger, S. Lienhard, A. Sturm
C. Unger
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M. Hardegger
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S. Lienhard
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A. Sturm
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Published April 1994. DOI: https://doi.org/10.1104/pp.104.4.1351

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Abstract

Carrot (Daucus carota), like most other plants, contains various isoenzymes of acid [beta]-fructofuranosidase ([beta]F) (invertase), which either accumulate as soluble polypeptides in the vacuole (isoenzymes I and II) or are ionically bound to the cell wall (extracellular [beta]F). Using antibodies against isoenzyme I of carrot soluble [beta]F, we isolated several cDNA clones encoding polypeptides with sequences characteristic of [beta]Fs, from bacteria, yeast, and plants. The cDNA-derived polypeptide of one of the clones contains all partial peptide sequences of the purified isoenzyme I and thus codes for soluble acid [beta]F isoenzyme I. A second clone codes for a related polypeptide (63% identity and 77% similarity) with characteristics of isoenzyme II. These two soluble [beta]Fs, have acidic isoelectric points (3.8 and 5.7, respectively) clearly different from the extracellular enzyme, which has a basic isoelectric point of 9.9. Marked differences among the three nucleotide sequences as well as different hybridization patterns on genomic DNA gel blots prove that these three isoenzymes of carrot acid [beta]F are encoded by different genes and do not originate from differential splicing of a common gene, as is the case in the yeast Saccharomyces cerevisiae. All three carrot acid [beta]Fs, are preproenzymes with signal peptides and N-terminal propeptides. A comparison of the sequences of the soluble enzymes with the sequence of the extracellular protein identified C-terminal extensions with short hydrophobic amino acid stretches that may contain the information for vacuolar targeting.

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cDNA Cloning of Carrot (Daucus carota) Soluble Acid [beta]-Fructofuranosidases and Comparison with the Cell Wall Isoenzyme
C. Unger, M. Hardegger, S. Lienhard, A. Sturm
Plant Physiology Apr 1994, 104 (4) 1351-1357; DOI: 10.1104/pp.104.4.1351

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cDNA Cloning of Carrot (Daucus carota) Soluble Acid [beta]-Fructofuranosidases and Comparison with the Cell Wall Isoenzyme
C. Unger, M. Hardegger, S. Lienhard, A. Sturm
Plant Physiology Apr 1994, 104 (4) 1351-1357; DOI: 10.1104/pp.104.4.1351
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Plant Physiology
Vol. 104, Issue 4
Apr 1994
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  • Cloning of a Temperature-Regulated Gene Encoding a Chloroplast [omega]-3 Desaturase from Arabidopsis thaliana
  • Structural Features of the Maize sus1 Gene and Protein
  • Genes Galore: A Summary of Methods for Accessing Results from Large-Scale Partial Sequencing of Anonymous Arabidopsis cDNA Clones
Show more Molecular Biology and Gene Regulation

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