Isolation of mesocarp-specific cDNA clone from the oil palm via RT-PCR

Nurniwalis Abdul Wahab and Siti Nor Akmar Abdullah and Tan, Siang Hee (2003) Isolation of mesocarp-specific cDNA clone from the oil palm via RT-PCR. [Proceedings Paper]

Storage oil is produced in oil palm mesocarp. This oil contains 44% of palmitic acid, 39% of oleic acid, 10% of linoleic acid, less than 5% of stearic acid and a small percentage of myristic acid, linolenic acid, lauric acid and palmitoleic acid. During fruit development, oil accumulation starts at about 15 week after ant hes is (WAA) and stops when the fruit ripens at 20 WAA. Gene expression studies during this period is of interest to serve as background information to genetically engineer the oil palm in order to expand its applicability. To ensure the accumulation of the genetically engineered products are targeted to the mesocarp, promoter sequences that are responsible to direct the expression of the desired gene must be identified. The most important step towards isolating these essential mesocarp-specific promoters is isolation of mesocarp-specific genes. In this study, RT-PCR is chosen to isolate a cDNA clone strongly expressed in the mesocarp during the period of oil synthesis. Partial mesocarp-specific cDNAs generated via RT-PCR was successfully transformed into TA TOPO cloning vector and this was confirmed by restriction analysis. Identity search via internet showed 80% identity with score bit of 250 and E value of 2e-63 to genes with known identity in other monocot such as Oryza sativa. Preliminary result using Dot Blot analysis showed that the gene is highly expressed in the oil palm mesocarp. No expression was detected in leaf and kernel at 12 and 13 WAA. Efforts are currently focused on confirming the temporal and tissue-specificity of the isolated gene via Northern blot analysis. The full length clone will also be isolated via cDNA library screening and 5'RACE.

Full text available from:

Abstract

Storage oil is produced in oil palm mesocarp. This oil contains 44% of palmitic acid, 39% of oleic acid, 10% of linoleic acid, less than 5% of stearic acid and a small percentage of myristic acid, linolenic acid, lauric acid and palmitoleic acid. During fruit development, oil accumulation starts at about 15 week after ant hes is (WAA) and stops when the fruit ripens at 20 WAA. Gene expression studies during this period is of interest to serve as background information to genetically engineer the oil palm in order to expand its applicability. To ensure the accumulation of the genetically engineered products are targeted to the mesocarp, promoter sequences that are responsible to direct the expression of the desired gene must be identified. The most important step towards isolating these essential mesocarp-specific promoters is isolation of mesocarp-specific genes. In this study, RT-PCR is chosen to isolate a cDNA clone strongly expressed in the mesocarp during the period of oil synthesis. Partial mesocarp-specific cDNAs generated via RT-PCR was successfully transformed into TA TOPO cloning vector and this was confirmed by restriction analysis. Identity search via internet showed 80% identity with score bit of 250 and E value of 2e-63 to genes with known identity in other monocot such as Oryza sativa. Preliminary result using Dot Blot analysis showed that the gene is highly expressed in the oil palm mesocarp. No expression was detected in leaf and kernel at 12 and 13 WAA. Efforts are currently focused on confirming the temporal and tissue-specificity of the isolated gene via Northern blot analysis. The full length clone will also be isolated via cDNA library screening and 5'RACE.

[error in script]

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:	mesocarp
AGROVOC Term:	gene expression
AGROVOC Term:	plant genetics
AGROVOC Term:	polymerase chain reaction > polymerase chain reaction Prefer using PCRPCR
AGROVOC Term:	data analysis
AGROVOC Term:	enzymes
AGROVOC Term:	DNA cloning
AGROVOC Term:	genetic markers
Geographical Term:	Malaysia
Depositing User:	Nor Hasnita Abdul Samat
Date Deposited:	16 Jul 2024 03:11
Last Modified:	17 Jul 2024 04:44
URI:	http://webagris.upm.edu.my/id/eprint/839

Actions (login required)

View Item