The low-molecular-weight glutenin subunit proteins of primitive wheats. III. The genes from D-genome species

M. Ciaffi, Y. K. Lee, L. Tamas, R. Gupta, J. Skerritt, R. Appels

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Abstract

The isolation and characterisation by DNA sequencing of two different low molecular weight glutenin subunit (LMW-GS) genes from a genomic library derived from Triticum tauschii is described. These genes are similar (more than 90% similarity) but not identical to previously published LMW-GS gene sequences from cultivated wheats. A comparison of nucleotide sequence of the coding regions revealed the presence of insertions and deletions preferentially located in the region encoding the domains in the LMW-GS proteins rich in proline and glutamine and the middle part of the C-domain. The signal sequences, the amino-terminus and the remaining parts of the C-domain were conserved between all the LMW-GSs compared. The differences detected between the deduced amino-acid sequences in these three regions are only due to single nucleotide substitutions. The most important characteristic of all compared LMW-GS genes is the conservation of eight cysteine residues that could be involved in potential secondary or tertiary structure and disulphide-bond interactions. Comparisons between the 5' and 3' non-coding sequences of one of the isolated clones (LMW-16/10) with those of different prolamin genes from wheat, barley and rye led to the distinction of five different gene families, and confirmed the evolutionary relationships determined previously for these genes mainly on the basis of the coding region. In particular, the LMW-GS sequences are more closely related to the B-hordein sequences than to any other prolamin genes from wheat, barley and rye. Formal proof that the isolated genes coded for LMW-GSs, as defined by gel electrophoresis, was obtained by moving one of these genes (LMW-16/10) into a bacterial expression vector based on bacteriophage T7 RNA polymerase. The resulting plasmid directed the synthesis of large amounts of the mature form of the subunit in Escherichia coli. This protein exhibited solubility characteristics identical to those of the LMW-GSs and cross-reacted with antibodies reactive with these proteins.

Original languageEnglish
Pages (from-to)135-148
Number of pages14
JournalTheoretical and Applied Genetics
Volume98
Issue number1
DOIs
Publication statusPublished - Jan 1 1999

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Keywords

  • Bacterial expression
  • LMW-GS genes
  • Molecular evolution
  • Seed storage proteins
  • Triticum tauschii
  • Wheat quality

ASJC Scopus subject areas

  • Biotechnology
  • Agronomy and Crop Science
  • Genetics

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