Analysis of the oil palm palmitoyl-acyl carrier protein thioesterase gene expression in the transgenic Arabidopsis thaliana plants

Nurhafizah R., . and Abrizah O., . and Mohamad Arif A.M., . and Zaini A., . (2007) Analysis of the oil palm palmitoyl-acyl carrier protein thioesterase gene expression in the transgenic Arabidopsis thaliana plants. [Proceedings Paper]

This paper will discuss further the analysis of the thioesterase gene expression in the transgenic Arabidopsis thaliana plants. To investigate the affect of over expression of an oil palm thioesterase gene on fatty acid profiles an expression construct pCB35S was produced. In an attempt to elucidate the thioesterase in vivo specificity as well as test the efficiency of the sense constructs i.e. targeting thioesterase over expression to the cytoplasm and chloroplast the binary vector pCB35S was constructed which carried a full-length cDNA encoding palmitoyl-ACP thioesterase from oil palm EgcFatB and placed under the control of the cauliflower mosaic virus 35S promoter. The acyl-ACP thioesterase from oil palm was previously shown to exhibit in vitro hydrolytic activity for long chain acyl-ACPs. The plasmid was used to transform Arabidopsis thaliana plants by the Agrobacterium tumefaciens floral dip method. Selection of the transgenic plants was carried out using Basta as a selective marker via spray technique on soil. A total of 69 transgenic T3-plant lines were obtained. PCR assay was carried out for the transgenes TJ of pCB35S using gene specific primers AEP 1 and AC02 and the expected band of 750 bp was observed.

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Abstract

This paper will discuss further the analysis of the thioesterase gene expression in the transgenic Arabidopsis thaliana plants. To investigate the affect of over expression of an oil palm thioesterase gene on fatty acid profiles an expression construct pCB35S was produced. In an attempt to elucidate the thioesterase in vivo specificity as well as test the efficiency of the sense constructs i.e. targeting thioesterase over expression to the cytoplasm and chloroplast the binary vector pCB35S was constructed which carried a full-length cDNA encoding palmitoyl-ACP thioesterase from oil palm EgcFatB and placed under the control of the cauliflower mosaic virus 35S promoter. The acyl-ACP thioesterase from oil palm was previously shown to exhibit in vitro hydrolytic activity for long chain acyl-ACPs. The plasmid was used to transform Arabidopsis thaliana plants by the Agrobacterium tumefaciens floral dip method. Selection of the transgenic plants was carried out using Basta as a selective marker via spray technique on soil. A total of 69 transgenic T3-plant lines were obtained. PCR assay was carried out for the transgenes TJ of pCB35S using gene specific primers AEP 1 and AC02 and the expected band of 750 bp was observed.

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Item Type:	Proceedings Paper
AGROVOC Term:	ELAEIS GUINEENSIS
AGROVOC Term:	ARABIDOPSIS THALIANA
AGROVOC Term:	OIL PALMS
AGROVOC Term:	PCR
AGROVOC Term:	DNA
AGROVOC Term:	TRANSGENIC PLANTS
AGROVOC Term:	TRANSGENICS
AGROVOC Term:	GENE EXPRESSION
AGROVOC Term:	MALAYSIA
Geographical Term:	MALAYSIA
Depositing User:	Ms. Norfaezah Khomsan
Last Modified:	24 Apr 2025 05:27
URI:	http://webagris.upm.edu.my/id/eprint/15720

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