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2012 | 34 | 2 |

Tytuł artykułu

Characterization of a hydroxyproline-rich glycoprotein in pearl millet and its differential expression in response to the downy mildew pathogen Sclerospora graminicola

Warianty tytułu

Języki publikacji

EN

Abstrakty

EN
A monoclonal antibody, JIM 20, derived against an extensin type of hydroxyproline-rich glycoprotein (HRGP) from pea, showed high affinity for HRGP in pearl millet [Pennisetum glaucum (L.) R. Br.]. Electrophoretic separation of Tris–SDS extracted proteins from suspension cells of pearl millet revealed a range of PMHRGP polypeptides having a glycan epitope, which reacted with JIM 20. A high molecular mass band, probably an HRGP aggregate or polymer, and a few low molecular mass polypeptides were recognized by JIM 20 during Western blot analysis. Treatment of pearl millet suspension cells with hydrogen peroxide in the presence of an endogenous peroxidase resulted in insolubilization of HRGP polypeptides with molecular weights between 45 and 33 kDa. To investigate the gene coding for an extensin type of HRGP, a fosmid-based genomic library of pearl millet having a fourfold genome coverage was constructed. A partial sequence of 378 bp of an HRGP gene was obtained by PCR amplification of pearl millet DNA with a primer pair designed from the conserved regions of monocotyledon extensin type of HRGPs. Screening the genomic library using the homologous probe developed from the 378-bp PCR product resulted in the isolation of five fosmid clones. Restriction mapping of these fosmids resulted in an 11.8-kb region around an HRGP gene in pearl millet. The newly characterized gene, PM-HRGP, had all the characteristic features of a monocotyledon extensin type of HRGP. An intron at the 30 untranslated region of the gene was identified by cDNA cloning. Differential expression of the PM-HRGP gene was observed during compatible and incompatible interactions of pearl millet with the downy mildew pathogen Sclerospora graminicola (Sacc) Schroet. Induced expression of the gene was observed only in case of an incompatible interaction.

Słowa kluczowe

Wydawca

-

Rocznik

Tom

34

Numer

2

Opis fizyczny

p.779-791,fig.,ref.

Twórcy

autor
  • Molecular Biology of Plants, Groningen Biomolecular Sciences and Biotechnology Institute, Centre for Life Sciences, University of Groningen, Nijenborgh 7, 9747 AG Groningen, The Netherlands
  • Department of Studies in Biotechnology, Manasagangotri, University of Mysore, Mysore 570006, Karnataka, India
autor
  • Department of Studies in Biotechnology, Manasagangotri, University of Mysore, Mysore 570006, Karnataka, India
  • Department of Studies in Biotechnology, Manasagangotri, University of Mysore, Mysore 570006, Karnataka, India
autor
  • Sylphium Life Sciences, Duurswoldlaan 22, 9727 DJ Groningen, The Netherlands
autor
  • Department of Studies in Biotechnology, Manasagangotri, University of Mysore, Mysore 570006, Karnataka, India
autor
  • Molecular Biology of Plants, Groningen Biomolecular Sciences and Biotechnology Institute, Centre for Life Sciences, University of Groningen, Nijenborgh 7, 9747 AG Groningen, The Netherlands

Bibliografia

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Bibliografia

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