Identification and molecular characterization of alcohol dehydrogenase (ADH) and its promoter region in sago palm (Metroxylon sagu)

Sago palm (Metroxylon sagu) is one of the main economic crops for the state of Sarawak. It is one of the few plant species that thrive on peat swamp area. The waterlogged condition in such areas causes oxygen deprivation to the root part and triggers the anaerobic pathway. Molecular analysis on sa...

Full description

Saved in:

Bibliographic Details
Main Author:	Yasotha, A/P Sundaraj
Format:	Thesis
Language:	English
Published:	2008
Subjects:	TP Chemical technology
Online Access:	http://ir.unimas.my/id/eprint/28818/2/Yasotha%28fulltext%29.pdf
Tags:	Add Tag No Tags, Be the first to tag this record!

id	my-unimas-ir.28818
record_format	uketd_dc
spelling	my-unimas-ir.288182023-06-08T06:54:13Z Identification and molecular characterization of alcohol dehydrogenase (ADH) and its promoter region in sago palm (Metroxylon sagu) 2008 Yasotha, A/P Sundaraj TP Chemical technology Sago palm (Metroxylon sagu) is one of the main economic crops for the state of Sarawak. It is one of the few plant species that thrive on peat swamp area. The waterlogged condition in such areas causes oxygen deprivation to the root part and triggers the anaerobic pathway. Molecular analysis on sago helps in understanding the ability of the palm to tolerate anaerobic conditions. In this study, proteins from various parts of the sago palm such as the roots (dry and submerged) and leaves were extracted successfully. The proteins were subjected for analysis on agarose and polyacralymide gel, and the presence of alcohol dehydrogenase (ADH), lactate dehydrogenase (LDH) as well as superoxide dismutase (SOD) enzymes was detected through specific staining. The results indicated that ADH is present in all analyzed sago tissues and three variants were detected based on migration on gel. However, the banding patterns and the intensity varied among the tissues with the highest expression of ADH and SOD enzymes being reported in the sago shoots. Apart from that, three putative Adh genes from sago palm denoted as M I, M2 and M3 were isolated through Polymerase Chain Reaction (PCR) method using specific primers in this study. Based on the multiple alignment analysis, the three loci are found to be highly diverged and contain a common 23 bp nucleotide which is conserved in other Adh plant species. Phylogeny analysis indicated that the Adh loci of palm (Arecaceae) are segregated from that of other plant Adh families such as from the grass family (poaceae) and Rosaceae, corresponding to the findings of prevIous phylogenetic analysis. All these results concluded that three unique putative Adh genes from sago palm have been successfully isolated. One sago palm Adh gene, referred to here as M2 was used as template for primer designing for DNA walking to further isolate the upstream region which regulates the transcription of the particular gene. The DNA Walking technique has been adapted since genomic library construction yielded a low number of titer, 1.0 x 105 pfulml. Through DNA Walking, a promoter region was successfully identified. Manual and computational analysis of the promoter region revealed the presence of important elements such as TAT A box, CCAA T box and other putative regulatory elements at their respective locations. Universiti Malaysia Sarawak (UNIMAS) 2008 Thesis http://ir.unimas.my/id/eprint/28818/ http://ir.unimas.my/id/eprint/28818/2/Yasotha%28fulltext%29.pdf text en validuser masters Universiti Malaysia Sarawak (UNIMAS) Faculty of Resource Science and Technology
institution	Universiti Malaysia Sarawak
collection	UNIMAS Institutional Repository
language	English
topic	TP Chemical technology
spellingShingle	TP Chemical technology Yasotha, A/P Sundaraj Identification and molecular characterization of alcohol dehydrogenase (ADH) and its promoter region in sago palm (Metroxylon sagu)
description	Sago palm (Metroxylon sagu) is one of the main economic crops for the state of Sarawak. It is one of the few plant species that thrive on peat swamp area. The waterlogged condition in such areas causes oxygen deprivation to the root part and triggers the anaerobic pathway. Molecular analysis on sago helps in understanding the ability of the palm to tolerate anaerobic conditions. In this study, proteins from various parts of the sago palm such as the roots (dry and submerged) and leaves were extracted successfully. The proteins were subjected for analysis on agarose and polyacralymide gel, and the presence of alcohol dehydrogenase (ADH), lactate dehydrogenase (LDH) as well as superoxide dismutase (SOD) enzymes was detected through specific staining. The results indicated that ADH is present in all analyzed sago tissues and three variants were detected based on migration on gel. However, the banding patterns and the intensity varied among the tissues with the highest expression of ADH and SOD enzymes being reported in the sago shoots. Apart from that, three putative Adh genes from sago palm denoted as M I, M2 and M3 were isolated through Polymerase Chain Reaction (PCR) method using specific primers in this study. Based on the multiple alignment analysis, the three loci are found to be highly diverged and contain a common 23 bp nucleotide which is conserved in other Adh plant species. Phylogeny analysis indicated that the Adh loci of palm (Arecaceae) are segregated from that of other plant Adh families such as from the grass family (poaceae) and Rosaceae, corresponding to the findings of prevIous phylogenetic analysis. All these results concluded that three unique putative Adh genes from sago palm have been successfully isolated. One sago palm Adh gene, referred to here as M2 was used as template for primer designing for DNA walking to further isolate the upstream region which regulates the transcription of the particular gene. The DNA Walking technique has been adapted since genomic library construction yielded a low number of titer, 1.0 x 105 pfulml. Through DNA Walking, a promoter region was successfully identified. Manual and computational analysis of the promoter region revealed the presence of important elements such as TAT A box, CCAA T box and other putative regulatory elements at their respective locations.
format	Thesis
qualification_level	Master's degree
author	Yasotha, A/P Sundaraj
author_facet	Yasotha, A/P Sundaraj
author_sort	Yasotha, A/P Sundaraj
title	Identification and molecular characterization of alcohol dehydrogenase (ADH) and its promoter region in sago palm (Metroxylon sagu)
title_short	Identification and molecular characterization of alcohol dehydrogenase (ADH) and its promoter region in sago palm (Metroxylon sagu)
title_full	Identification and molecular characterization of alcohol dehydrogenase (ADH) and its promoter region in sago palm (Metroxylon sagu)
title_fullStr	Identification and molecular characterization of alcohol dehydrogenase (ADH) and its promoter region in sago palm (Metroxylon sagu)
title_full_unstemmed	Identification and molecular characterization of alcohol dehydrogenase (ADH) and its promoter region in sago palm (Metroxylon sagu)
title_sort	identification and molecular characterization of alcohol dehydrogenase (adh) and its promoter region in sago palm (metroxylon sagu)
granting_institution	Universiti Malaysia Sarawak (UNIMAS)
granting_department	Faculty of Resource Science and Technology
publishDate	2008
url	http://ir.unimas.my/id/eprint/28818/2/Yasotha%28fulltext%29.pdf
_version_	1783728363987271680

Identification and molecular characterization of alcohol dehydrogenase (ADH) and its promoter region in sago palm (Metroxylon sagu)

Similar Items