%0 Journal Article
%T Isolation of Alcohol Dehydrogenase cDNA and Basal Regulatory Region from Metroxylon sagu
%A Ching Ching Wee
%A Hairul Azman Roslan
%J ISRN Molecular Biology
%D 2012
%R 10.5402/2012/839427
%X Alcohol dehydrogenase (Adh) is a versatile enzyme involved in many biochemical pathways in plants such as in germination and stress tolerance. Sago palm is plant with much importance to the state of Sarawak as one of the most important crops that bring revenue with the advantage of being able to withstand various biotic and abiotic stresses such as heat, pathogens, and water logging. Here we report the isolation of sago palm Adh cDNA and its putative promoter region via the use of rapid amplification of cDNA ends (RACE) and genomic walking. The isolated cDNA was characterized and determined to be 1464£¿bp long encoding for 380 amino acids. BLAST analysis showed that the Adh is similar to the Adh1 group with 91% and 85% homology with Elaeis guineensis and Washingtonia robusta, respectively. The putative basal msAdh1 regulatory region was further determined to contain promoter signals of TATA and AGGA boxes and predicted amino acids analyses showed several Adh-specific motifs such as the two zinc-binding domains that bind to the adenosine ribose of the coenzyme and binding to alcohol substrate. A phylogenetic tree was also constructed using the predicted amino acid showed clear separation of Adh from bacteria and clustered within the plant Adh group. 1. Introduction Alcohol dehydrogenase (Adh) is an enzyme involved in various biological activities such as in the germination and abiotic stresses in plants [1¨C3]. Previous studies have shown that there are between two or three Adh loci in flowering plants with exception in Arabidopsis [4, 5]. Previous Adh protein work on sago palm, a flood-tolerant plant, by Roslan et al. [6] detected the presence of Adh in the leaf and roots. A higher Adh enzyme expression was observed in sago palm young shoots compared to the other part of Metroxylon sagu [6]. The finding was consistent with those of Padmanabhan and Sahi [7] that reported a greater increase in Adh activity in the leaves than the roots of sunflower that was treated with high phosphorus. In contrast, in flood-intolerant plants such as Arabidopsis and pea, increased Adh activity was determined in the roots than in the shoots under anaerobic condition [8, 9]. A higher expression level in different tissue and developmental stage may be because the cells are dividing and exposed to many stresses [10]. The discovery of Adh protein expression in young leaf prompted the work to isolate the Adh gene from sago palm. The isolation of the regulatory region was also conducted to further understand the regulation of Adh in sago palm. Adh gene have been isolated from
%U http://www.hindawi.com/journals/isrn.molecular.biology/2012/839427/