Alamsyah, Firman and Widyasari, Wiwit Budi and Suhandono, Sony (2020) Characterization of pyrophosphate-dependent phosphofructokinase α-subunit gene from sugarcane showing gene without intron. IOP Conference Series: Earth and Environmental Science, 457 (1). 012078. ISSN 1755-1315
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Abstract
Pyrophosphate-dependent Phosphofructokinase or PFP is an enzyme that regulate sucrose metabolism. It consists of α- and β-subunits, which encoded by PFPα and PFPβ genes, respectively. Sugarcane PFPa has a strong function in glycolysis and has the potency to be engineered to increase sugarcane yield. Hence, the purpose of this work was to isolate, clone and characterize the sugarcane PFPα gene. Total RNA was isolated from leafrolls of TD 91 sugarcane variety. cDNA synthesis followed by DNA amplification of PFPα gene were performed using degenerate primers. cDNA and DNA fragments were ligated into pGEM-T Easy vector, which were subsequently introduced to E. coli competent cells. EcoRI were used to cut the plasmids for sequencing. Finally, homology searching was conducted using BLASTn, and then the nucleotide sequence was translated to a protein sequence using Bioedit. The results showed that PFPα cDNA fragment was 900 bp in length. The translated PFPa protein showed binding sites for fructose-6-phosphate and fructose-1,6-biphosphate, which are conserved in all family members of PFP. In silico analysis of the DNA fragment showed gene without intron. In conclusion, the PFPα gene from sugarcane has been successfully isolated, cloned and characterized.
Item Type: | Article |
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Additional Information: | This article is indexed in Q3 Journal Ranking Index, after presented in The 3rd International Conference on Biosciences held on 8 August 2019 at IPB International Convention Centre, Bogor - Indonesia; TURNITIN's similarity index result was checked based on rules stated in "Pedoman Operasional Penilaian Angka Kredit Kenaikan Jabatan Akademik/Pangkat Dosen" (http://lldikti12.ristekdikti.go.id/wp-content/uploads/2019/03/PO-PAK-2019_MULAI-BERLAKU-APRIL-2019.pdf) pp. 23-25 |
Uncontrolled Keywords: | sugarcane, PFPa gene, sugar metabolic engineering, escherichia coli, amino acid sequence, gene, Saccharum officinarum, protein, nucleotide sequence, phosphofructokinase, binding site, subunit, plasmid, clone, cDNA, intron, vector (genetics) |
Subjects: | 500 Natural sciences & mathematics > 500 Science 500 Natural sciences & mathematics > 580 Plants (Botany) |
Divisions: | Universitas Al-Azhar Indonesia (UAI) > Fakultas Sains dan Teknologi (FST) > Biologi |
Depositing User: | Rifda Jilan |
Date Deposited: | 22 Sep 2021 02:43 |
Last Modified: | 22 Nov 2021 10:52 |
URI: | http://eprints.uai.ac.id/id/eprint/1708 |
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