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Riza A PUTRANTO
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Agriculture, Biological Sciences & Forestry

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Isolation and characterization of Dehydrin promoter region from sugarcane (Saccharum officinarum L.) Hayati MINARSIH; Sonny SUHANDONO; Anissa K FUADI; Tati KRISTIANTI; Riza A PUTRANTO; Deden SUKMAJDAYA; . SUSTIPRIJATNO
E-Journal Menara Perkebunan Vol 88, No 1 (2020): April, 2020
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (867.853 KB) | DOI: 10.22302/iribb.jur.mp.v88i1.363

Abstract

The development of molecular biology techniques nowadays has enabled to engineer drought tolerant sugarcane by genetic engineering to accelerate the breeding program. Dehydrin (DHN) is known to have an important role in plant response and adaptation to abiotic stresses (drought, high salinity, cold, heat, etc.). While plant tissues are subjected to drought stress (dehydration), DHN protein is accumulated to high content throughout all vegetative or generative tissues. The research aimed to isolate and characterize the DHN promoter from sugarcane that can be used as transformation material in generating drought tolerant sugarcane. Specific primers for DHN promoter amplification were designed and DHN promoter region was successfully isolated by PCR cloning method. Two putative promoter sequences were identified namely Pr-1DHNSo and Pr-2DHNSo. In silicoanalyses were carried out and cis-regulatory elements motifs that play a role in adaptation on abiotic stress as well as biotic stress including ABRE, MBS, CGTCA-motif, TGACG-motif, GARE-motif, P-box TCA-element and Box-W1 were identified. The promoter Pr-1DHNSo was then cloned into pBI121 expression vector by Overlap Extention PCR (OE-PCR) for further characterization. Functional test of the promoter construct pBI- Pr-1DHNSo was conducted through Agrobacterium transformation into sugarcane calli. GUS assay and PCR analysis showed that the DHN promoter was transformed and expressed in the sugarcane calli.
Karakterisasi gen penyandi lipase dari kapang Rhizopus oryzae dan Absidia corymbifera Characterization of gene encoding lipase from fungus Rhizopus oryzae and Absidia corymbifera Riza A PUTRANTO; Djoko SANTOSO; . TRI-PANJI; . SUHARYANTO; Asmini BUDIANI
E-Journal Menara Perkebunan Vol 74, No 1: Juni 2006
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (471.091 KB) | DOI: 10.22302/iribb.jur.mp.v74i1.118

Abstract

SummaryLipase is a group of enzymes which catalyze fat hydrolysis. Lipase is recently used to produce diacylglycerol (DAG) from triacylglycerol (TAG). Lipase  can be used to produce healthy oil. Having a rich biodiversity, Indonesia has the opportunity to produce lipase using indigenous microbes, such as molds. This research aimed to detect  LIPASE  gene on several strains of molds employing PCR technique. Genomic DNAs were isolated from four strains of molds (M. sitophila, R. oryzae, R. microsporus, and A. corymbifera). Heterologous primers for LIPASE  were designed based on the conserved region of 12 LIPASE  sequences accessed from GenBank and used to amplify the genomic DNA resulted in a 466 bp fragmen. BLAST analysis showed that the bands of DNAs have high homology with common lipase protein in several strains of  Rhizopus.Ringkasan Lipase merupakan kelompok enzim yang berfungsi sebagai biokatalis hidrolisis lemak. Lipase banyak digunakan untuk konversi triasilgliserol (TAG) menjadi diasilgliserol (DAG). Penggunaan lipase penting untuk produksi minyak sehat (healthy oil). Indonesia dengan keanekaragaman hayati tinggi berpeluang besar   mengembangkan   produksi   lipase   dari mikroba lokal, salah satunya adalah kapang. Deteksi gen merupakan langkah awal dalam upaya peningkatan produksi lipase melalui rekayasa genetika. DNA genomik empat galur kapang (M. sitophila, R. oryzae, R. microsporus, dan A. corymbifera) telah berhasil diisolasi. Sepasang primer heterologous telah berhasil dirancang berdasarkan daerah terkonservasi 12 sekuen gen LIPASE dari GenBank. Amplikon DNA yang diperoleh pada PCR menggunakan pasangan primer RLP memiliki panjang 466 bp. Analisis BLAST memperlihatkan bahwa amplikon PCR memiliki homologi yang tinggi dengan protein LIPASE  beberapa galur Rhizopus. 
Karakterisasi gen penyandi lipase dari kapang Rhizopus oryzae dan Absidia corymbifera Characterization of gene encoding lipase from fungus Rhizopus oryzae and Absidia corymbifera Riza A PUTRANTO; Djoko SANTOSO; . TRI-PANJI; . SUHARYANTO; Asmini BUDIANI
Menara Perkebunan Vol. 74 No. 1: 74 (1), 2006
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v74i1.118

Abstract

SummaryLipase is a group of enzymes which catalyze fat hydrolysis. Lipase is recently used to produce diacylglycerol (DAG) from triacylglycerol (TAG). Lipase  can be used to produce healthy oil. Having a rich biodiversity, Indonesia has the opportunity to produce lipase using indigenous microbes, such as molds. This research aimed to detect  LIPASE  gene on several strains of molds employing PCR technique. Genomic DNAs were isolated from four strains of molds (M. sitophila, R. oryzae, R. microsporus, and A. corymbifera). Heterologous primers for LIPASE  were designed based on the conserved region of 12 LIPASE  sequences accessed from GenBank and used to amplify the genomic DNA resulted in a 466 bp fragmen. BLAST analysis showed that the bands of DNAs have high homology with common lipase protein in several strains of  Rhizopus.Ringkasan Lipase merupakan kelompok enzim yang berfungsi sebagai biokatalis hidrolisis lemak. Lipase banyak digunakan untuk konversi triasilgliserol (TAG) menjadi diasilgliserol (DAG). Penggunaan lipase penting untuk produksi minyak sehat (healthy oil). Indonesia dengan keanekaragaman hayati tinggi berpeluang besar   mengembangkan   produksi   lipase   dari mikroba lokal, salah satunya adalah kapang. Deteksi gen merupakan langkah awal dalam upaya peningkatan produksi lipase melalui rekayasa genetika. DNA genomik empat galur kapang (M. sitophila, R. oryzae, R. microsporus, dan A. corymbifera) telah berhasil diisolasi. Sepasang primer heterologous telah berhasil dirancang berdasarkan daerah terkonservasi 12 sekuen gen LIPASE dari GenBank. Amplikon DNA yang diperoleh pada PCR menggunakan pasangan primer RLP memiliki panjang 466 bp. Analisis BLAST memperlihatkan bahwa amplikon PCR memiliki homologi yang tinggi dengan protein LIPASE  beberapa galur Rhizopus. 
Isolation and characterization of Dehydrin promoter region from sugarcane (Saccharum officinarum L.) Hayati MINARSIH; Sonny SUHANDONO; Anissa K FUADI; Tati KRISTIANTI; Riza A PUTRANTO; Deden SUKMAJDAYA; . SUSTIPRIJATNO
Menara Perkebunan Vol. 88 No. 1 (2020): 88 (1), 2020
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v88i1.363

Abstract

The development of molecular biology techniques nowadays has enabled to engineer drought tolerant sugarcane by genetic engineering to accelerate the breeding program. Dehydrin (DHN) is known to have an important role in plant response and adaptation to abiotic stresses (drought, high salinity, cold, heat, etc.). While plant tissues are subjected to drought stress (dehydration), DHN protein is accumulated to high content throughout all vegetative or generative tissues. The research aimed to isolate and characterize the DHN promoter from sugarcane that can be used as transformation material in generating drought tolerant sugarcane. Specific primers for DHN promoter amplification were designed and DHN promoter region was successfully isolated by PCR cloning method. Two putative promoter sequences were identified namely Pr-1DHNSo and Pr-2DHNSo. In silicoanalyses were carried out and cis-regulatory elements motifs that play a role in adaptation on abiotic stress as well as biotic stress including ABRE, MBS, CGTCA-motif, TGACG-motif, GARE-motif, P-box TCA-element and Box-W1 were identified. The promoter Pr-1DHNSo was then cloned into pBI121 expression vector by Overlap Extention PCR (OE-PCR) for further characterization. Functional test of the promoter construct pBI- Pr-1DHNSo was conducted through Agrobacterium transformation into sugarcane calli. GUS assay and PCR analysis showed that the DHN promoter was transformed and expressed in the sugarcane calli.
Co-Authors . SUSTIPRIJATNO Anissa K FUADI Asmini Budiani Deden SUKMAJDAYA Djoko Santoso Hanny Hafiar Hayati MINARSIH Sonny SUHANDONO Suharyanto TATI KRISTIANTI Tri Panji