Molecular and General Genetics MGG

, Volume 245, Issue 2, pp 177–186 | Cite as

Structural characterization and promoter activity analysis of the γ-kafirin gene from sorghum

  • Fernando A. de Freitas
  • José Andrés Yunes
  • Marcio J. da Silva
  • Paulo Arruda
  • Adilson Leite
Original Paper


A genomic clone encoding the γ-kafirin gene from sorghum was isolated and sequenced. A 2938 bp sequenced fragment includes an intronless open reading frame of 636 nucleotides encoding a putative polypeptide of 212 amino acids. Comparison of the deduced amino acid sequence of γ-kafirin with the published sequences of γ-prolamins of maize, and Coix revealed highly conserved domains. The N-terminal region of these proteins contains the conserved hexapeptide PPPVHL, which is repeated eight times in γ-zein, four times in γ-kafirin and three times in γ-coixin. The number of PPPVHL repeats accounts predominantly for the differences in the molecular weights of γ-prolamins. Several putative regulatory sequences common to the γ-kafirin and γ-zein genes were identified in both the 5′ and the 3′ flanking regions. Putative GCN4-like regulatory sequences were found at positions −192 and −476 in the 5′ flanking region of γ-kafirin. In the 3′ noncoding region, three putative polyadenylation signals, two AATAAT and one AATGAA, were found at positions + 658, + 716, and + 785, respectively. In order to investigate the role of the putative GCN4-like motifs and other possible cis-acting element(s) of the γ-kafirin promoter, a series of deleted and chimeric promoter constructs were introduced into maize, Coix and sorghum tissues by particle bombardment. Histochemical analysis of β-glucuronidase (GUS) activity in different tissues indicated that the element(s) responsible for tissue specificity is probably located in the 285-bp proximal region of the promoter, while the remaining promoter sequence seems to carry the element(s) responsible for the quantitative response.

Key words

Gene expression Tissue specificity Seed storage protein Sorghum γ-Kafirin 


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Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • Fernando A. de Freitas
    • 1
  • José Andrés Yunes
    • 1
  • Marcio J. da Silva
    • 1
  • Paulo Arruda
    • 1
    • 2
  • Adilson Leite
    • 1
  1. 1.Centro de Biologia Molecular e Engenharia GenéticaUniversidade Estadual de Campinas, Cidade Universitária “Zeferino Vaz”CampinasBrazil
  2. 2.Departamento de Genética e Evolução, 1BUniversidade Estadual de CampinasCampinasBarzil

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