Molecular cloning of pea cDNA encoding cycloartenol synthase and its functional expression in yeast

Biol Pharm Bull. 1997 Jul;20(7):770-5. doi: 10.1248/bpb.20.770.

Abstract

The cDNA encoding cycloartenol synthase [EC 5.4.99.8] has been isolated from pea seedling by an efficient PCR using sets of degenerate primers based on the highly conserved sequences of the known 2,3-oxidosqualene cyclase cDNAs. The obtained cDNA contains a 2271-bp open reading frame and is encoding a predicted protein of 757 amino acids with high homology (81%) to Arabidopsis thaliana cycloartenol synthase. The PCR-amplified open reading frame (ORF) has been inserted into pYES2, an expression vector in yeast, under the control of galactose-inducible promoter. Significant cycloartenol synthase activity has been found in the homogenate of the yeast transformed with the plasmid containing PCR-amplified ORF.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • DNA, Complementary
  • Intramolecular Transferases*
  • Isomerases / genetics*
  • Molecular Sequence Data
  • Open Reading Frames
  • Pisum sativum / enzymology
  • Pisum sativum / genetics*
  • Saccharomyces cerevisiae / genetics*
  • Sequence Homology, Amino Acid

Substances

  • DNA, Complementary
  • Isomerases
  • Intramolecular Transferases
  • cycloartenol synthase

Associated data

  • GENBANK/D89619