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PMID: 8980496 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Molecular characterization of transketolase (EC 2.2.1.1) active in the Calvin cycle of spinach chloroplasts.

Plant molecular biology ·Vol. 32 ·No. 3 ·1996-11-00 ·Pages 475-84

Flechner A, Dressen U, Westhoff P, Henze K, Schnarrenberger C, Martin W

Abstract

A cDNA encoding the Calvin cycle enzyme transketolase (TKL; EC 2.2.1.1) was isolated from Sorghum bicolor via subtractive differential hybridization, and used to isolate several full-length cDNA clones for this enzyme from spinach. Functional identity of the encoded mature subunit was shown by an 8.6-fold increase of TKL activity upon induction of Escherichia coli cells that overexpress the spinach TKL subunit under the control of the bacteriophage T7 promoter. Chloroplast localization of the cloned enzyme is shown by processing of the in vitro synthesized precursor upon uptake by isolated chloroplasts. Southern blot-analysis suggests that TKL is encoded by a single gene in the spinach genome. TKL proteins of both higher-plant chloroplasts and the cytosol of non-photosynthetic eukaryotes are found to be unexpectedly similar to eubacterial homologues, suggesting a possible eubacterial origin of these nuclear genes. Chloroplast TKL is the last of the demonstrably chloroplast-localized Calvin cycle enzymes to have been cloned and thus completes the isolation of gene probes for all enzymes of the pathway in higher plants.

MeSH Terms
Amino Acid Sequence Base Sequence Chloroplasts/enzymology,genetics Cloning, Molecular Escherichia coli/genetics Genes, Plant/genetics Molecular Sequence Data Molecular Weight Photosynthesis/genetics Phylogeny Protein Precursors/metabolism Recombinant Fusion Proteins Sequence Analysis, DNA Spinacia oleracea/enzymology,genetics Transketolase/chemistry,genetics,metabolism
Chemicals
Protein Precursors Recombinant Fusion Proteins Transketolase
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Flechner A
Institut für Pflanzenphysiologie, Berlin, Germany.
Dressen U
Westhoff P
Henze K
Schnarrenberger C
Martin W
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Article Info
Journal
Plant molecular biology
Abbr.
Plant Mol Biol
ISSN
0167-4412
DOI
10.1007/BF00019099
Published
1996-11-00
Pages
475-84
Language
English
Region
Netherlands
NLM ID
9106343
Subset
IM
Databases
GENBANK
H55032, L76554
External Links
PubMed source DOI: 10.1007/BF00019099 View journal details
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