Cloning and expression in Escherichia coli of a cDNA coding for the oleoyl-acyl carrier protein thioesterase from coriander (Coriandrum sativum L.).
A cDNA for the oleoyl-acyl carrier protein (ACP) thioesterase (E.C. 3.1.2.14) from coriander seed endosperm (Coriandrum sativum) was isolated using a safflower oleoyl-ACP thioesterase cDNA probe. The coriander cDNA coded for a 42.3 kDa protein including a putative 40 amino acid plastid targeting transit peptide. The gene was expressed in Escherichia coli and the recombinant protein was isolated to homogeneity by alkyl-ACP affinity and anion-exchange chromatography. The pure protein showed a high thioesterase activity for oleoyl-ACP vs. other acyl-ACPs and therefore was identified as the coriander oleoyl-ACP thioesterase. Antibodies were raised against the recombinant protein and used to detect the coriander thioesterase in enriched endosperm fractions.
Dörmann
P
P
Department of Botany and Plant Pathology, Michigan State University, East Lansing 48824.
Kridl
J C
JC
Ohlrogge
J B
JB
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004926:Escherichia coli / Q000235:genetics*; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D017365:Spices; D013869:Thiolester Hydrolases / Q000235:genetics*
D018076:DNA, Complementary; D013869:Thiolester Hydrolases; C018748:oleoyl-(acyl-carrier-protein) hydrolase