Molecular cloning of pea cDNA encoding cycloartenol synthase and its functional expression in yeast.
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.
Morita
M
M
Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.
Shibuya
M
M
Lee
M S
MS
Sankawa
U
U
Ebizuka
Y
Y
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D019751:Intramolecular Transferases; D007535:Isomerases / Q000235:genetics*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D018532:Pisum sativum / Q000201:enzymology / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D007535:Isomerases; D019751:Intramolecular Transferases; C021923:cycloartenol synthase