Isolation and characterization of sago palm alcohol dehydrogenase promoter region

Nurul Fazlin, Binti Abdul Rahman. (2007) Isolation and characterization of sago palm alcohol dehydrogenase promoter region. [Final Year Project Report] (Unpublished)

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Abstract

Alcohol dehydrogenase (.4dh) gene is very essential in plant as it is responsible to help plant to adapt with environmental stress, especially when deprivation of oxygen is occurred. In order to have the gene to be expressed, promoter plays an important role to synthesis the protein encoded by Adh gene. Thus, this study was carried out to amplifY the Adh gene from Sago and consequent to isolate the Adh promoter region using genome walking strategy. eTAB method was used to extract high quality of DNA from sago. The DNA was cu t into smaller fragment using hexacutters restriction enzyme and the 3 'overhangs was ligated with overhang primer. Following ligation, 5'end overhangs are formed on the DNA, which serves sites for adaptor primers and nested primers fur peR amplification in combination with the gene-specific primers. Multiple bands were obtained in secondary DNA walking peR. These four bands were excised from gel and sent for direct sequencing. From the four samples, only one sample obtained a good sequencing result. The sequence was analyzed using software to find the promoter region.

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