Molecular cloning and characterization of gene encoding gibberellin 2-oxidase (GA20X) from shorea parvifolia dyer parvifolia

Ong, Li Yew. (2006) Molecular cloning and characterization of gene encoding gibberellin 2-oxidase (GA20X) from shorea parvifolia dyer parvifolia. [Final Year Project Report] (Unpublished)

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Abstract

Semidwarf tree varieties have provided many advantages to forestry as produce higher wood quality. The gene which is responsible for dwarfism varieties is the gibberellin 2-oxidase (GA20x). The catabolic pathway of the gibberellin (GA) in plant which involve the GA20x enzyme will catalyses the conversion of bioactive GAs compound into inactive form that cause dwarfism on plant. In this study, Shorea parvifi)lia Dyer parvifolia was chosen due to its commercial value and strong adaptability. The RNA was isolated from the species using the optimized RNA isolation protocol for Shorea parvljOlia Dyer parvifolia from Plant Genomic Laboratory, Unimas. Two pairs of primers, namely At primer and Hypp primer were constructed based on the known GA20x sequences obtained from gene bank databases. Reverse transcriptase-Polymerase Chain Reaction (RT-PCR) was performed and a DNA fragment of 515 bp from the PCR amplification with the At primer was purified. The DNA fragment was cloned into pGEMT®-Easy vector and sent for sequencing. However, the sequence result showed low homology to Bacillus anthracis, Homo sapiens chromosome 17, Human BAC clone CTA-20803, Uncultured bacterium clone YC01A01, and Helicobacter pylori of unknown function instead ofGA20x.

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