cDNA cloning and expression analysis of tocopherol cyclase gene from various oil palm tissues (Elaeis guineensis and Elaeis oleifera)

Tocopherol cyclase (TC) which catalyses the formation of γ-tocopherol from reduced (quinol form) 2,3-dimethyl-6-phytyl-1,4-benzoquinone (DMPBQ) is one of the key enzymes in tocopherol biosynthetic pathway. The aim of this study is isolate the cDNA clone encoding TC and to study its expression profil...

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Main Author:	Lai, Mun Seong
Format:	Thesis
Language:	English English
Published:	2011
Subjects:	Molecular cloning Vitamin E Oil palm - Analysis
Online Access:	http://psasir.upm.edu.my/id/eprint/27007/1/FP%202011%2032R.pdf
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id	my-upm-ir.27007
record_format	uketd_dc
spelling	my-upm-ir.270072022-01-26T05:49:08Z cDNA cloning and expression analysis of tocopherol cyclase gene from various oil palm tissues (Elaeis guineensis and Elaeis oleifera) 2011 Lai, Mun Seong Tocopherol cyclase (TC) which catalyses the formation of γ-tocopherol from reduced (quinol form) 2,3-dimethyl-6-phytyl-1,4-benzoquinone (DMPBQ) is one of the key enzymes in tocopherol biosynthetic pathway. The aim of this study is isolate the cDNA clone encoding TC and to study its expression profile in different oil palm tissues and different developmental stages in the mesocarp. A partial cDNA 0.5 kb in length was amplified using gene-specific primer from an oil palm expressed sequence tags sequence and found to have high sequence identity with other plant species TC such as Oryza sativa (81%) and Helianthus annus (76%) at the amino acid level. Rapid amplification of cDNA ends (RACE) and end to end PCR successfully amplified the cDNA for the full mature protein of TC from two oil palm species (E. guineensis and E. oleifera) of 1. 233 kb in length encoding 411 amino acids. The amino acid identity between the E. guineensis and E. oleifera sequence was 99.6%. Both sequences have three long stretches of hydrophobic residues and 30 carboxylterminal amino acid unique for plants (not found in cynobacteria TC). However, the reported conserved 5 amino acid carboxyl domain KPPGL in plant TC has 1 variant amino acid in both oil palm sequences (RPPGL). Quantitative gene expression analysis performed using Real-time PCR showed expression of E. guineensis TC in mesocarp was moderately high at 7 w.a.a., dropped at 10 w.a.a then increased steadily from 10 w.a.a and reached a maximum level at 17 w.a.a and the expression level dropped significantly at 19 w.a.a. The overall expression levels in leaves were lower than mesocarp with young leaves having a higher expression level than mature leaves. Real-time gene copy number analysis indicated only one copy of TC exists in the oil palm genome. The isolated mature protein sequence of oil palm TC can be used in functional study to understand the vitamin E biosynthetic process in oil palm and plants in general. It can also be used in genetic engineering efforts to modify vitamin E content and composition in plants. Molecular cloning Vitamin E Oil palm - Analysis 2011 Thesis http://psasir.upm.edu.my/id/eprint/27007/ http://psasir.upm.edu.my/id/eprint/27007/1/FP%202011%2032R.pdf application/pdf en public masters Universiti Putra Malaysia Molecular cloning Vitamin E Oil palm - Analysis Faculty of Agriculture English
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English English
topic	Molecular cloning Vitamin E Oil palm - Analysis
spellingShingle	Molecular cloning Vitamin E Oil palm - Analysis Lai, Mun Seong cDNA cloning and expression analysis of tocopherol cyclase gene from various oil palm tissues (Elaeis guineensis and Elaeis oleifera)
description	Tocopherol cyclase (TC) which catalyses the formation of γ-tocopherol from reduced (quinol form) 2,3-dimethyl-6-phytyl-1,4-benzoquinone (DMPBQ) is one of the key enzymes in tocopherol biosynthetic pathway. The aim of this study is isolate the cDNA clone encoding TC and to study its expression profile in different oil palm tissues and different developmental stages in the mesocarp. A partial cDNA 0.5 kb in length was amplified using gene-specific primer from an oil palm expressed sequence tags sequence and found to have high sequence identity with other plant species TC such as Oryza sativa (81%) and Helianthus annus (76%) at the amino acid level. Rapid amplification of cDNA ends (RACE) and end to end PCR successfully amplified the cDNA for the full mature protein of TC from two oil palm species (E. guineensis and E. oleifera) of 1. 233 kb in length encoding 411 amino acids. The amino acid identity between the E. guineensis and E. oleifera sequence was 99.6%. Both sequences have three long stretches of hydrophobic residues and 30 carboxylterminal amino acid unique for plants (not found in cynobacteria TC). However, the reported conserved 5 amino acid carboxyl domain KPPGL in plant TC has 1 variant amino acid in both oil palm sequences (RPPGL). Quantitative gene expression analysis performed using Real-time PCR showed expression of E. guineensis TC in mesocarp was moderately high at 7 w.a.a., dropped at 10 w.a.a then increased steadily from 10 w.a.a and reached a maximum level at 17 w.a.a and the expression level dropped significantly at 19 w.a.a. The overall expression levels in leaves were lower than mesocarp with young leaves having a higher expression level than mature leaves. Real-time gene copy number analysis indicated only one copy of TC exists in the oil palm genome. The isolated mature protein sequence of oil palm TC can be used in functional study to understand the vitamin E biosynthetic process in oil palm and plants in general. It can also be used in genetic engineering efforts to modify vitamin E content and composition in plants.
format	Thesis
qualification_level	Master's degree
author	Lai, Mun Seong
author_facet	Lai, Mun Seong
author_sort	Lai, Mun Seong
title	cDNA cloning and expression analysis of tocopherol cyclase gene from various oil palm tissues (Elaeis guineensis and Elaeis oleifera)
title_short	cDNA cloning and expression analysis of tocopherol cyclase gene from various oil palm tissues (Elaeis guineensis and Elaeis oleifera)
title_full	cDNA cloning and expression analysis of tocopherol cyclase gene from various oil palm tissues (Elaeis guineensis and Elaeis oleifera)
title_fullStr	cDNA cloning and expression analysis of tocopherol cyclase gene from various oil palm tissues (Elaeis guineensis and Elaeis oleifera)
title_full_unstemmed	cDNA cloning and expression analysis of tocopherol cyclase gene from various oil palm tissues (Elaeis guineensis and Elaeis oleifera)
title_sort	cdna cloning and expression analysis of tocopherol cyclase gene from various oil palm tissues (elaeis guineensis and elaeis oleifera)
granting_institution	Universiti Putra Malaysia
granting_department	Faculty of Agriculture
publishDate	2011
url	http://psasir.upm.edu.my/id/eprint/27007/1/FP%202011%2032R.pdf
_version_	1747811570580193280

cDNA cloning and expression analysis of tocopherol cyclase gene from various oil palm tissues (Elaeis guineensis and Elaeis oleifera)

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