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All Journal HAYATI Journal of Biosciences Farmasains : Jurnal Farmasi dan Ilmu Kesehatan Journal of Tropical Life Science : International Journal of Theoretical, Experimental, and Applied Life Sciences Jurnal Kedokteran Brawijaya The Journal of Experimental Life Sciences (JELS) Biotropika Research Journal of Life Science Natural B Edu Geography Jurnal Pengembangan Teknologi Informasi dan Ilmu Komputer Jurnal Penelitian Pendidikan IPA (JPPIPA) Indonesian Journal of Cardiology Buletin Anatomi dan Fisiologi (Bulletin of Anatomy and Physiology) Care : Jurnal Ilmiah Ilmu Kesehatan Jurnal Jamu Indonesia Nusantara Science and Technology Proceedings Berkala Penelitian Hayati Makara Journal of Science Indonesian Journal of Jamu
Sri Widyarti
Biology Department, Faculty Of Mathematics And Natural Sciences Brawijaya University, Malang Indonesia
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Journal : Makara Journal of Science

 1 
Employing SecA Recombinant Protein to Generate Polyclonal Antibodies for the Rapid Identification of Phytoplasma Fatinah, Arik Arubil; Rifa'i, Muhaimin; Arumingtyas, Estri Laras; Widyarti, Sri
Makara Journal of Science Vol. 29, No. 3
Publisher : UI Scholars Hub

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Abstract

Phytoplasma, a plant pathogen that threatens plant health, can induce diseases such as yellow wilt, virescence, phyllody, and witches’ broom. Although early detection methods using polyclonal and monoclonal antibodies have been developed, synthetic peptides offer advantages such as higher titers, predictable antigenic characteristics and superior sensitivity and specificity. Epitope mapping plays a key role in designing peptide antigens and includes gene synthesis, transformation, expression assessment, large-scale production, and purification. A study using New Zealand male White rabbits was conducted to generate antibodies against phytoplasma. The SecA (395–470) sequence, obtained from the preprotein translocase meets the criteria of being non-homologous, nonallergic, and antigenic. The three-dimensional structure of SecA (395–470) has a sequential similarity of 77–97% with certain phytoplasma types, and the protein purity exceeds 90%. The production of polyclonal antibodies was successful, achieving titers greater than 1:512,000. The SWISS model was used to predict the 3D structure of SecA in phytoplasma species, revealing structural homology with other phytoplasma species. The recombinant protein antigen SecA was able to induce high-titer antibody formation (>1:512.000). The developed lateral flow immunoassay (LFA) detects phytoplasma in plants using purified and conjugated antibodies. The peptide design, derived from Aster yellows witches-broom (SrI), effectively detects phytoplasma from various groups, especially Cactus witches’ broom phytoplasma (SrII-C) and Peanut witches’ broom phytoplasma (SrII-A). The SrI group phytoplasma was chosen as a reference sequence due to its` ability to infect plants across the broadest host range. LFIA was tested on samples from both phytoplasma-infected and healthy plants using nested PCR. The SecA sequence was successfully produced and used as an immunogen candidate against phytoplasmas.
Co-Authors A'liyatur Rosidah A.A. Ketut Agung Cahyawan W A’liyatur Rosyidah Aditya Tri Hernowo Ahmad Lubab Ahmad Soni Ahmad Soni Amandia Dewi Permana Shita Apik Budi Santoso Arief Azhary Aris Soewondo Aris Suwondo Aristyani, Sherry Askandar Tjokroprawiro Attabik Mukhammad Amrillah Aulanni'am Azzamuddin, Haidar Bella Novinda Cholilia Abadiatul Masruroh Cholilia Abadiatul Masruroh Damayanti, Febriane Eka Daniar Kusumawati Dayu Nirwana Putri Dewi Parlina Ningrum Dian Siswanto Djanggan Sargowo Dyah Kinasih Wuragil Eko Puji Astuti Erly Nur Aisyah Estri Laras Arumingtyas Farida Rachmawati, Farida Fatinah, Arik Arubil Hafiedz Maulana Harits Amrulloh Intan Sartika Is Karima Jannah, Nur Karima, Is Khansa, Shafanony Khoirin Maghfiroh Linda Kartika Dewi Lulut Dwi Nurmamulyosari M. Irsyad Nur Maftuch Maftuch Marji Marji Moch Sasmito Djati Mochammad Pratama Viadi Mohammad Mabrur Muhaimin Rifa'i Muhaimin Rifa'i Muhaimin Rifa'i Muhaimin Rifa'i Muhaimin Rifa'i Muhaimin Rifa'i Muhaimin Rifai'i Muhamad Aris Widodo Naqiyah A. Mulachelah Nashi Widodo Natasya Adiba Zahrah Noer Hasanah Nunung Harijati Nur Permatasari Nur Permatasari Nuzulul Hikmah Olly Indrajani Puspitoputri, Alya H. Radita Intan Aisyah Pratiwi Rasjad Indra Retno Susilowati Retty Ratnawati Rifa'i, Muhaimin Rifa'i, Muhaimin Rifa’i, Muhaimin Rike Wahyuningroom Rike Wahyuningroom Ririn Rochmawati Rismawati, Ria Rizki Amalia Rochmatika, Lailiyavina Rodiyati Azrianingsih Rodiyati Azrianingsih Rosidah, A'liyatur Rosyidah, A’liyatur Sakti, Sefihara Paramitha Sapti Puspitarini Sari, Fikriya Novita Satuman Satuman Serafinah Indriyani Sherry Aristyani Siti Imama Khoiriyah Sofy Permana Soni, Ahmad SRI RAHAYU Sri Rahayu Sri Rahayu Sri Rahayu Sri Rahayu Sri Rahayu Suroso Suroso Sutiman B. Sumitro Sutiman B. Sumitro Sutiman B. Sumitro Sutiman B. Sumitro Sutiman Bambang Sumitro Suwondo, Aris Syahputra Wibowo Teguh Suprihatin Teguh Suprihatin Ulinnuha, Maria Wibi Riawan Widodo Widodo Yayu Tsamrotul Fuadah, Yayu Tsamrotul Yoga Dwi Jatmiko YOGA DWI JATMIKO Yuni Kilawati Zauhani Kusnul