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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
1996-11-5
pubmed:databankReference
pubmed:abstractText
A molecular biological study on limonene synthase that catalyzes the cyclization of geranyldiphosphate to yield the olefin 4(S)-limonene, an intermediate in the biosynthesis of a monoterpenoid, perillaldehyde, in Perilla frutescens Britton has been carried out. We isolated and characterized 10 cDNAs homologous to spearmint limonene synthase cDNA from an expression library constructed from cotyledons of a Perilla strain homozygous for two pairs of dominant genes, G and H, which are responsible for the formation of 4(S)-limonene. Two of these cDNA clones were functionally expressed in Escherichia coli, yielding enzymes which were catalytically active in generating 4(S)-limonene from geranyldiphosphate. The longest open reading frame in the representative cDNA clone PFLC1 consisted of 1812 nucleotides corresponding to 603 amino acids. Its identity to the ORFs of spearmint limonene synthase, tobacco 5-epi-aristolochene synthase, and castor bean casbene synthase were 65, 35, and 30%, respectively. Genomic Southern blot analyses of various genotypes (GGHH, GGhh, ggHH, and gghh) of P. frutescens suggested that more than one copy of the PFLC1 DNA exists in strains having the HH genotype. In contrast, no PFLC1 DNA sequences were found in the genomes of strains with the hh genotype that are incapable of producing cyclohexanoid monoterpenes for lack of limonene synthase activity. Northern blot analysis, using a PFLC1 3'-flanking region as a hybridization probe, showed that PFLC1 mRNA accumulated in all the aerial parts of the GGHH plants, particularly in the leaves. In the ggHH plants, on the other hand, PFLC1 mRNA was detected only in minute amounts in the stem and calyx.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0003-9861
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
332
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
280-7
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:8806736-Amino Acid Sequence, pubmed-meshheading:8806736-Base Sequence, pubmed-meshheading:8806736-Cloning, Molecular, pubmed-meshheading:8806736-DNA, Complementary, pubmed-meshheading:8806736-DNA, Plant, pubmed-meshheading:8806736-Escherichia coli, pubmed-meshheading:8806736-Gas Chromatography-Mass Spectrometry, pubmed-meshheading:8806736-Gene Expression, pubmed-meshheading:8806736-Genes, Plant, pubmed-meshheading:8806736-Intramolecular Lyases, pubmed-meshheading:8806736-Isomerases, pubmed-meshheading:8806736-Molecular Sequence Data, pubmed-meshheading:8806736-Plants, Edible, pubmed-meshheading:8806736-Plants, Medicinal, pubmed-meshheading:8806736-RNA, Messenger, pubmed-meshheading:8806736-Recombinant Proteins, pubmed-meshheading:8806736-Restriction Mapping, pubmed-meshheading:8806736-Sequence Homology, Amino Acid, pubmed-meshheading:8806736-Tissue Distribution
pubmed:year
1996
pubmed:articleTitle
cDNA cloning, characterization, and functional expression of 4S-(-)-limonene synthase from Perilla frutescens.
pubmed:affiliation
Faculty of Pharmaceutical Sciences, Kyoto University, Japan.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't