Isolation of partial DNA sequence coding for Lycopene β-Cyclase from oil palm (E. guineensis)

O. A. Rasid and R. Singh and K. Harikrishna and Ho, C. L. and Cheah, S. C. (2003) Isolation of partial DNA sequence coding for Lycopene β-Cyclase from oil palm (E. guineensis). [Proceedings Paper]

The uses and value of plant minor components such as carotenoids, flavonoid and sterols have recently been discovered. Carotenoids in particular have been shown to be potential for its anticancer and antioxidant activities. Extensive studies have been conducted to elucidate the plant carotenoid pathway. Oil palm is known to be the richest natural source for carotene. However, to date, there has been no work carried out to elucidate the pathway in this species. This paper is the first report on the isolation of cDNA clones coding for lycopene B-cyclase from oil palm and most importantly the first effort towards understanding the oil palm carotenoid biosynthesis pathway. In this work, we have successfully obtained a 675 bp fragment through RT-PCR. The fragment was cloned into PCRII-TOPO vector and its complete DNA sequence was obtained. DNA sequence analysis of the representative clones showed that the clones are highly identical to other lycopene β-cyclases at about 80% identity. Primers were then designed based on the DNA sequence to facilitate the amplification of the 3' and 5' end regions of the genes. A 662 bp fragment corresponding to the 3'-end region of the gene was successfully amplified, cloned and sequenced. A consensus sequence of 1277 bp was generated from the two fragments (i.e. middle and 3'-end regions) including a poly A-tail. A partial open reading frame (ORF) of 1149 bp was identified by comparison of its deduced amino acid sequence. The ORF codes for 382 amino acid residues and lacks about 100 amino acid residues of its 5'-end region. Comparison to other published sequence lycopene β-cyclases, oil palm lycopene β-cyclase was shown to share high identity at both DNA and amino acid levels.

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Abstract

The uses and value of plant minor components such as carotenoids, flavonoid and sterols have recently been discovered. Carotenoids in particular have been shown to be potential for its anticancer and antioxidant activities. Extensive studies have been conducted to elucidate the plant carotenoid pathway. Oil palm is known to be the richest natural source for carotene. However, to date, there has been no work carried out to elucidate the pathway in this species. This paper is the first report on the isolation of cDNA clones coding for lycopene B-cyclase from oil palm and most importantly the first effort towards understanding the oil palm carotenoid biosynthesis pathway. In this work, we have successfully obtained a 675 bp fragment through RT-PCR. The fragment was cloned into PCRII-TOPO vector and its complete DNA sequence was obtained. DNA sequence analysis of the representative clones showed that the clones are highly identical to other lycopene β-cyclases at about 80% identity. Primers were then designed based on the DNA sequence to facilitate the amplification of the 3' and 5' end regions of the genes. A 662 bp fragment corresponding to the 3'-end region of the gene was successfully amplified, cloned and sequenced. A consensus sequence of 1277 bp was generated from the two fragments (i.e. middle and 3'-end regions) including a poly A-tail. A partial open reading frame (ORF) of 1149 bp was identified by comparison of its deduced amino acid sequence. The ORF codes for 382 amino acid residues and lacks about 100 amino acid residues of its 5'-end region. Comparison to other published sequence lycopene β-cyclases, oil palm lycopene β-cyclase was shown to share high identity at both DNA and amino acid levels.

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Item Type:	Proceedings Paper
Additional Information:	Available at Perpustakaan Sultan Abdul Samad, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia. TP684 P3I61 2003 Call Number
AGROVOC Term:	oil palm > oil palm Prefer using Elaeis guineensisElaeis guineensis
AGROVOC Term:	lycopene
AGROVOC Term:	molecular biology
AGROVOC Term:	gene cloning
AGROVOC Term:	multilocus sequence analysis
AGROVOC Term:	researchers > researchers Prefer using scientistsscientists
AGROVOC Term:	genetic resources
AGROVOC Term:	DNA sequences
Geographical Term:	Malaysia
Depositing User:	Nor Hasnita Abdul Samat
Date Deposited:	23 Jul 2024 02:01
Last Modified:	23 Jul 2024 02:01
URI:	http://webagris.upm.edu.my/id/eprint/841

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