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

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.

• Authors: M Morita, M Shibuya, M S Lee, U Sankawa, Y Ebizuka
• Journal: Biological & pharmaceutical bulletin (Biol Pharm Bull) Vol. 20 Issue 7 Pg. 770-5 (Jul 1997) ISSN: 0918-6158 [Print] Japan
• PMID: 9255418 (Publication Type: Journal Article)

Chemical References

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

Topics

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