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International Journal of Pure & Applied Bioscience (IJPAB)
Year : 2018, Volume : 6, Issue : 3
First page : (1) Last page : (10)
Article doi: : http://dx.doi.org/10.18782/2320-7051.6566

Isolation, Characterization and Gene Sequencing of Partial Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) Gene of Diospyros discolor Willd.

Pineda, Adrian Renz R.¹*, Acuña, Edsel Ivan F.² and Paitan, Virginia P.³
College of Science, Bulacan State University, City of Malolos, Bulacan, Philippines
*Corresponding Author E-mail: adrianrenzpineda10@gmail.com
Received: 15.05.2018 | Revised: 21.06.2018 | Accepted: 27.06.2018  

 ABSTRACT

The Diospyros discolor Willd. is an endemic and vulnerable plant species in the Philippines known for its premium kamagong timber and mabolo fruit that possesses nutritional, medicinal and pharmacological properties. However, due to its limited genetic information, it is less likely be used to gene expression studies. This study worked for the initial step – isolation of a specific housekeeping gene, glycerlaldehyde-3-phosphate dehydrogenase.  The gDNA from the leaf tissue was extracted using CTAB method, then subjected it to gradient PCR in order to amplify the GAPDH gene in an optimized temperature using the designed primers. Samples K2 and L1 at 57.4oC and 57.8oC were purified and sequenced by the Macrogen, Korea. Results of in silico analyses indicated that the isolated gene is partial GAPDH gene that contains 687 base pairs with an exact reading frame of 300 base pairs which corresponds to 100 amino acids. The conserved domains from the amino acid sequence proved that it belongs to the GAPDH NAD binding domain superfamily and GAPDH C-terminal domain superfamily. The relative plant species was determined using BLASTn and BLASTp where Clematoclethra scandens, Acrostichum aureum, Vittaria graminifolia and Antrophyum latifolium showed the highest relevance with 85% identity. The sequence data were submitted and accepted by GenBank, NCBI with an accession number of MH234383.1. The success of this study enables addition to the  genetic background of D. discolor Willd. that can be employed to its gene expression analysis and of other endemic and/or endangered related flora.

Key words: partial GAPDH gene, Diospyros discolor, genomic DNA, Gradient PCR

Full Text : PDF; Journal doi : http://dx.doi.org/10.18782

Cite this article: Pineda, A.R.R. and Acuña, E.I.F., Paitan, V.P., Isolation, Characterization and Gene Sequencing of Partial Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) Gene of Diospyros discolor Willd., Int. J. Pure App. Biosci.6(3): 1-10 (2018). doi: http://dx.doi.org/10.18782/2320-7051.6566