Expression, cloning, and immunological analysis of buckwheat (Fagopyrum esculentum Moench) seed storage proteins

J Agric Food Chem. 2001 Apr;49(4):1825-9. doi: 10.1021/jf0011485.

Abstract

cDNA of buckwheat (Fagopyrum esculentum Moench) was isolated from immature seeds harvested 14 days after pollination. Two genes, designated FA02 and FA18, were found to encode legumin-like proteins and were expressed during seed development. The deduced amino acid sequence of FA02 was identical to the N-terminal amino acid domain of BW24KD, which was believed to be a major buckwheat allergen (Urisu, A.; Kondo, Y.; Morita, Y.; Yagi, E.; Tsuruta, M.; Yasaki, T.; Yamada, K.; Kuzuya, H.; Suzuki, M.; Titani, K.; Kurosawa, K. Isolation and characterization of a major allergen in buckwheat seeds. In Current Advances in Buckwheat Research; Shinshu University Press: Matsumoto, Japan, 1995; pp 965--974). It was predicted that FA02 would be cleaved to generate two separate components, a 41.3 kDa alpha-subunit and a 21 kDa beta-subunit. Antiserum was raised against the deduced FA02 beta-subunit, and immunoblotting of total protein from buckwheat seeds (F. esculentum M. and Fagopyrum tartaricum Gaertn.) revealed that several groups of proteins reacted with the antiserum. Polypeptides in the 23--25 kDa range displayed the greatest reactivity.

MeSH terms

  • Allergens / genetics*
  • Allergens / immunology
  • Amino Acid Sequence
  • Cloning, Molecular
  • DNA, Complementary / chemistry
  • DNA, Complementary / isolation & purification
  • Electrophoresis, Agar Gel
  • Fagopyrum / chemistry*
  • Gene Expression Regulation, Plant
  • Gene Library
  • Immunoblotting
  • Molecular Sequence Data
  • Plant Proteins / chemistry
  • Plant Proteins / genetics*
  • Plant Proteins / immunology
  • Seeds / chemistry*

Substances

  • Allergens
  • DNA, Complementary
  • Plant Proteins

Associated data

  • GENBANK/D87980
  • GENBANK/D87982