Isolation and Characterization of the GA 20-Oxidase cDNA from Sago Palm (Metroxylon sagu Rottb.)

Bala, Jamel and Mohd. Hasnain, Hussain and Mohd. Azib, Salleh and Noraini, Busri (2011) Isolation and Characterization of the GA 20-Oxidase cDNA from Sago Palm (Metroxylon sagu Rottb.). Asia-Pacific Journal of Molecular Biology and Biotechnology, 19 (2). pp. 83-93. ISSN 0128-7451

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HETEROLOGOUS EXPRESSION OF XYLANASE GENE FROM KLEBSIELLA PNEUMONIAE IN E. COLI BL21 (DE3) FOR POTENTIAL USE IN GREEN TECHNOLOGY (abstract).pdf

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Abstract

GA 20-oxidase is involved in controlling stem elongation, maturity and flowering. Based on published conserved amino acid sequences of plant GA 20-oxidase cDNA clones, oligonucleotide primers were constructed and used to amplify partial sequence of the GA 20-oxidase gene of Metroxylon sagu. A 500 bp PCR product was obtained. BLAST analysis showed that the PCR product was homologous with the GA 20-oxidase gene from other plant species. The PCR product was labelled using the Digoxenin (DIG)-labelling system and used as a probe to obtain the full-length GA 20-oxidase gene sequence of a genomic fragment using the genome walking method. The genomic sequence was used for primer construction which were subsequently used to amplify a full-length cDNA copy of the GA 20-oxidase gene. The cDNA obtained was cloned into a pPCR-Script Amp SK (+) vector for sequence confirmation. The 1161 bp sequence obtained from the cDNA copy was compared with the genomic sequence of GA 20-oxidase. Comparison between genomic and cDNA fragments indicated that the GA 20-oxidase gene from sago palm is comprised of two introns and three exons. Heterologous expression of the GA 20-oxidase cDNA in Escherichia coli showed a similar expression pattern as the endogenous GA 20-oxidase of sago palm.

Item Type: Article
Uncontrolled Keywords: DNA sequencing, GA 20-Oxidase, Genome Walking, Sago palm, Southern hybridization, Western hybridization, unimas, university, universiti, Borneo, Malaysia, Sarawak, Kuching, Samarahan, ipta, education, research, Universiti Malaysia Sarawak
Subjects: S Agriculture > S Agriculture (General)
Divisions: Academic Faculties, Institutes and Centres > Faculty of Resource Science and Technology
Depositing User: Karen Kornalius
Date Deposited: 08 Dec 2015 06:31
Last Modified: 08 Dec 2015 06:31
URI: http://ir.unimas.my/id/eprint/9979

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