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.
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Conference Article|
December 01 2000
Cloning of a palmitoyl-acyl carrier protein thioesterase from oil palm
A. Othman;
A. Othman
1
1School of Biological Sciences, University of Bristol, Woodland Road, Bristol BS8 1UG, U.K.
1To whom correspondence should be addressed (e-mail abi.othman@bristol.ac.uk)
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C. Lazarus;
C. Lazarus
1School of Biological Sciences, University of Bristol, Woodland Road, Bristol BS8 1UG, U.K.
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T. Fraser;
T. Fraser
1School of Biological Sciences, University of Bristol, Woodland Road, Bristol BS8 1UG, U.K.
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K. Stobart
K. Stobart
1School of Biological Sciences, University of Bristol, Woodland Road, Bristol BS8 1UG, U.K.
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Publisher: Portland Press Ltd
Received:
June 30 2000
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© 2000 Biochemical Society
2000
Biochem Soc Trans (2000) 28 (6): 619–622.
Article history
Received:
June 30 2000
Citation
A. Othman, C. Lazarus, T. Fraser, K. Stobart; Cloning of a palmitoyl-acyl carrier protein thioesterase from oil palm. Biochem Soc Trans 1 December 2000; 28 (6): 619–622. doi: https://doi.org/10.1042/bst0280619
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