Genome walking of cysteine protease gene in Sago palm

Lee, Yen Sze (2012) Genome walking of cysteine protease gene in Sago palm. [Final Year Project Report] (Unpublished)

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Abstract

Sago palm or Metroxylon sagu is a monocotyledonous plant that belongs to the family Palmae and subfamily Calamoideae. In Sarawak, sago palm is one of the economically important agricultural crops. Cysteine protease plays a pivotal function in the growth and development of plant. It is also vital in programmed cell death (PCD). Cysteine protease gene has been known to be involved in regulated expression cysteine protease in sago palm. However, there are not much research have been carried out. Therefore, this study was performed to detect the regulatory elements such as promoter region for cysteine protease gene expression in sago palm. In this study, DNA was extracted by using a modified Doyle and Doyle method. Isolated DNA was cut into smaller fragments using restriction enzymes and the 3' overhangs were ligated to overhanging primers. The adaptor-ligated DNA was used to conduct genome walking. Bands were observed in the PCR amplification. However, the bands were too faint to be extracted from the gel and sent for sequencing though several PCR optimizations were done. Hence, the promoter region of cysteine protease gene was not obtained.

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