Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
Molecular Cloning of Pea cDNA Encoding Cycloartenol Synthase and Its Functional Expression in Yeast
Masayo MORITAMasaaki SHIBUYAMoon-Soon LEEUshio SANKAWAYutaka EBIZUKA
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JOURNALS FREE ACCESS

1997 Volume 20 Issue 7 Pages 770-775

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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.

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© The Pharmaceutical Society of Japan
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