Open AccessIsolation and functional analysis of a cDNA encoding a myrcene synthase from holm oak ( Quercus ilex L.)
Author(s) -
Fischbach Robert J.,
Zimmer Wolfgang,
Schnitzler JörgPeter
Publication year - 2001
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.1432-1033.2001.02519.x
Subject(s) - myrcene , monoterpene , biology , complementary dna , gene , atp synthase , heterologous expression , biochemistry , microbiology and biotechnology , botany , recombinant dna , essential oil , limonene
An 859‐bp cDNA segment of a terpene synthase gene was amplified by PCR from the evergreen sclerophyllous holm oak ( Quercus ilex L.) using heterologous primers for conserved regions of terpene synthase genes ( TPS ) in dicotyledonous plants. Based on the sequence of this segment, homologous primers were designed for amplification by RACE‐PCR of a cDNA segment carrying the monoterpene synthase gene myrS . The gene encodes a protein of 597 amino acids including an N‐terminal putative plastid transit peptide. The gene without the segment encoding the transit peptide was cloned by PCR into a bacterial expression vector. Expression in Escherichia coli yielded an active monoterpene synthase, which converted geranyl diphosphate (GDP) predominantly into the acyclic monoterpene myrcene and to a very small extent into cyclic monoterpenes. Sequence comparison with previously cloned monoterpene synthases revealed that the myrcene synthase from Q. ilex belongs to the TPSb subfamily.