Construction of plant transformation vector to contain metroxylon sagu alcohol dehydrogenase 1

Kon, Iris Chu Lin (2012) Construction of plant transformation vector to contain metroxylon sagu alcohol dehydrogenase 1. [Final Year Project Report] (Unpublished)

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Abstract

Alcohol dehydrogenase (ADH) is the critical enzyme responsible for the fermentative metabolism in plants especially under abiotic stress conditions. The levels of expression of Adh could be different in respective parts of plant in individual species. In this study, E. coli harbouring pET41(a)+msAdhl was used as the source to isolate the msAdhl gene through alkaline lysis method in order to be ligated into pGSAI131 to produce a plant transformation vector. The conserved region of msAdhl of 1.1 kb was amplified through PCR by using primer set of 5' Ndel-adh and 3'_msAdhl-Xhol. The msAdhl fragments with the concentration of 64.81 ng/Ill were restricted with Nde I and Xho I to remove the 3'-terminal adenines. Restriction digestion was performed on pGSAI131 at Nco I site to obtain linearized plasmid. The staggered ends of both the insert fragments and the linearized pGSAll31 were blunted by using T4 DNA Polymerase. The insert was ligated into pGSAll31 using T4 DNA Ligase before transformed into E. coli XLI Blue though heat-shock method. The verification of successfully constructed plasmids with correct orientation of msAdhl was performed via Colony PCR and sent for sequencing.

Item Type: Final Year Project Report
Additional Information: Project Report (B.Sc.) -- Universiti Malaysia Sarawak, 2012.
Uncontrolled Keywords: Alcohol, Biotechnology, Adhl, fermentative pathway, stress, open reading frame, recombinant plasmid, unimas, university, universiti, Borneo, Malaysia, Sarawak, Kuching, Samarahan, ipta, education, undergraduate, research, Universiti Malaysia Sarawak
Subjects: Q Science > Q Science (General)
Q Science > QK Botany
Divisions: Academic Faculties, Institutes and Centres > Faculty of Resource Science and Technology
Faculties, Institutes, Centres > Faculty of Resource Science and Technology
Depositing User: Karen Kornalius
Date Deposited: 01 Jul 2015 02:08
Last Modified: 11 Aug 2023 03:27
URI: http://ir.unimas.my/id/eprint/8127

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