Cloning of a palmitoyl-acyl carrier protein thioesterase from oil palm. (December 2000)

• Record Type: Journal Article
• Authors: Othman, A.; Lazarus, C.; Fraser, T.; Stobart, K.
• Is Part Of: Biochemical Society transactions. Volume 28:Number 6(2000)
• Page Start: 619
• Page End: 622
• Publication Date: 2000-12
• Subjects: cDNA -- genome -- palmitoyl-ACP thioesterase -- recombinant protein; ACP, acyl carrier protein -- IPTG, isopropyl β-D-thiogalactoside -- GST, glutathione S-transferase; Biochemistry -- Congresses; 572
• DOI: 10.1042/bst0280619
• Languages: English
• ISSNs: 0300-5127

Abstract

Abstract : A palmitoyl-acyl carrier protein (ACP) thioesterase cDNA clone was isolated from an oil palm cDNA library. The cDNA was expressed in Escherichia coli as a glutathione S-transferase fusion protein and a crude bacterial extract was assayed for acyl-CoA-hydrolysing activity. The recombinant enzyme was able to hydrolyse medium- and long-chain acyl-CoAs. Northernblot analysis showed a high level of gene expression in leaf, flower and 15-, 17- and 18-week mesocarp tissues. Low-level gene expression was detected in germinated seedlings and 8- and 12-week mesocarp tissues, but no transcript was detected in any kernel tissues. Southern-blot analysis indicated the presence of a single gene and we have also isolated a genomic clone using the cDNA as a probe. Two genomic fragments were subcloned and a 7 kb contiguous stretch of the oil palm genome was sequenced. Comparison of this sequence with the cDNA sequence identified a putative 93 amino acid transit peptide, most of which is missing from the cDNA. The coding region of the gene consisted of seven exons and six introns.