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Journal : Indonesian Journal of Biotechnology

Transient transformation of artemisinic aldehyde ∆ 11 (13) double bond reductase (dbr2) gene into Artemisia annua L. Elfahmi Elfahmi; Fany Mutia Cahyani; Andre Ditya Maulana Lubis; Tati Kristanti; Sony Suhandono
Indonesian Journal of Biotechnology Vol 21, No 2 (2016)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1326.152 KB) | DOI: 10.22146/ijbiotech.27956

Abstract

Global demand of antimalarial drug artemisinin has a gap with production capacity from existing sources since the low content of this compound from Artemia annua L. Genetic engineering-based strategy for A. annua plant on key enzymes in artemisinin biosynthetic pathway is needed. Artemisinic aldehyde ∆ 11 (13)  double bond reductase (dbr2) is one of the key enzyme on artemisinin biosynthesis which was studied in this research. Agrobacterium tumefaciens-mediated transformation of A. annua using dbr2 was carried out. Synthetic dbr2 was ligated into pCAMBIA1303 and transformed into Escherichia coli DH5α. pCAMBIA1303-dbr2 plasmid was transformed to A. tumefaciens AGL1. Leaves of  A. annua were infected by positive transformant of recombinant A. tumefaciens (OD600 ≈ 1) supplemented with acetosyringone 50 ppm, and Silwet S-408 0.02%. Samples were incubated in desiccators connected with vacuum pump, this method is called infiltration vacuum. Leaves were covered in dark for 45 min, and co-cultivated on MS co-cultivation media for 3 days. All leaves were washed in 300 ppm cefotaxime and divided into 2 parts; 3 leaves for GUS histochemical assay and 300 mg of leaves for HPLC analysis. Transient transformation was done in triplicate. In GUS histochemical assay, pCAMBIA1303 and pCAMBIA-dbr2 showed positive blue spot where coefficient of variance was less than 5%. PCR analysis for genomic DNA of transformed  A. annua showed a positive result of inserted dbr2 recombinant indicated by migration profile and direct sequencing analysis. It could be concluded that pCAMBIA-dbr2 construct and transformation into  A. annua have been successfully performed.
Co-Authors Adella Shindy Pratiwi Adji Mustiaji Agus Chahyadi Agus Chahyadi Agus Chahyadi Agus Supriadi Aisyah, Siti Zanuba Akbar Dana Akhirul Kahfi Syam Ana Indrayati Andi Rifki Rosandy Andre Ditya Maulana Lubis Andreanus A Soemardji Anggardiredja, Kusnandar Anita K Ari Sr Windyaswari Ari Sri Widyaswari Ari Sri Windyaswari Ari Sri Windyaswari, Ari Sri Ariranur Haniffadli ARTRI Astrid Indalifiany Ayu, Inna Puspa Bashari, Muhammad Hasan Beginer Subhan Budipramana, Krisyanti Choirul Huda Dana, Akbar DEBBIE SOEFIE RETNONINGRUM Desyane, Heipy Ketrin Dondy Arafat Edikresnha, Dhewa Fahrauk Faramayuda FAHRAUK FARAMAYUDA Fahrauk Faramayuda, Fahrauk Fany Mutia Cahyani Fatimah Arofiati Noor Frangky Sangande Ghozaly, Muchammad Reza Hapidin, Dian Ahmad Heipy Ketrin Desyane Jaka Permana Jenny Tri Yuspita Sari Khairunisa Harizqi Nurul Husna Khairunnisa Sy Khairurrijal khairurrijal Komar Ruslan Komar Ruslan Wirasutisna Komar Ruslan Wirasutisna Komar Ruslan Wirasutisna Kusnandar Anggardiredja Laode M.R. Al Muqarrabun Laode M.R. Al Muqarrabun Lusi Putri Dwita Luthfianti, Halida Rahmi Manalu, Meilina Marsinta Maria Immaculata Iwo Maria Immaculata Iwo Maria Immaculata Iwo Marlia Singgih Wibowo MARLIA SINGGIH WIBOWO Marlin Megalestin Raunsai Meilina Marsinta Manalu Melani, Putri Dwi Morawati, Soufni Muamar Abdillah Nabila K Nafisah, Nuraini Nebuchadnezzar Akbar NEVIATY PUTRI ZAMANI Ni Kadek Dita Cahyani, Ni Kadek Dita Niken Tunjung Murti Pratiwi Novriyandi Hanif Nurinanda Prisky Qomaladewi Nurul Nadiah Binti Abdul Rahman Oktiyas Muzaky Luthfi Pratiwi Ramadhani Feriza Rachmat Mauludin Ramelan, Riska Sigit RENALDI OKTAVIANUS Retno Wahyuningrum Retno Wahyuningrum Ridzka Magfirah Rika Amran Rika Hartati Riska Sigit Ramelan Rissyelly Rissyelly Rozida Khalid Sahidin Sahidin Salsabila, Syefira Sani, Lalu M. Iqbal Santoso, Winny Sartika Yuniarti Sawitri, Asti Shinta Maulida Rosandhy Sigid Pamungkas Wicaksono Silvy Julian Soemardji AA Sony Suhandono SONY SUHANDONO Sony Suhandono Soraya Riyanti Soraya Riyanti Sufyan Zainul Arifin Sukrasno Sukrasno Sukrasno Sukrasno Sukrasno Sukrasno Sukrasno Sukrasno Sukrasno Sulistiarini, Riski Sultan Salahudin Jamal Sumirtapura, Yeyet Cahyati Syafrizayanti, Syafrizayanti Syaikhul Aziz Tan MI Tati Kristanti TATI KRISTIANTI Tina Rostinawati Totik Sri Mariani Totik Sri Mariani Totik Sri Mariani Totik Sri Mariani Waresindo, William Xaveriano Weni Widy Astuti Winny Santoso Wirasutisna, Komar Ruslan Wirasutisna, Komar Ruslan Wiwin Herdwiani Yasa, Yanti Fitri Yeni Karlina Yodha, Agung Wibawa Mahatva Yolazenia Yolazenia Yosie Andriani Yuspita Sari, Jenny Tri Zaini Alfahmi