Analysis of Putative Sago palm cysteine protease cDNA open reading frame

Jong, Thin Thin (2013) Analysis of Putative Sago palm cysteine protease cDNA open reading frame. [Final Year Project Report] (Unpublished)

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Abstract

Cysteine protease is one of the types of proteolytic enzyme that present in Metroxylon sagu and has many roles in plant cells physiology and development process. In this study, the objective is to clone a putative cysteine protease gene derived from M. sagu into an expression vector. Firstly, the msCPR cDNA ORF which has been isolated from sago palm, was propagated in pBluescript plasmid. The clone was analyzed via restriction enzyme and PCR. The second objective is to clone the msCPR cDNA ORF into an expression vector, pET-41a(+). The last objective is to determine the correct insertion of the coding fragment of pET- 41a(+) which containing msCPR cDNA ORF through DNA sequencing reaction. A pBluescript vector containing the putative msCPR was checked via PCR and produced a fragment with the size of 750 bps. Then, the pET-41a(+) vector and msCPR cDNA ORF was successfully digested with Nde I and Not I restriction enzyme and subsequently ligated together. However, the determination of the correct insertion of the coding fragment of pET-41a(+)-msCPR cDNA ORF through DNA sequencing reaction was failed to be conducted.

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