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All Journal Jurnal Ilmu Pertanian Indonesia Jurnal Teknologi Dan Industri Pangan Jurnal Sain Veteriner Indonesian Journal of Tropical and Infectious Disease Indonesian Journal of Biotechnology UNEJ e-Proceeding SELAPARANG: Jurnal Pengabdian Masyarakat Berkemajuan Biosaintifika: Journal of Biology & Biology Education Jurnal Pengabdian Masyarakat Kesehatan Journal of Aquaculture and Fish Health Indian Journal of Forensic Medicine & Toxicology Berkala Penelitian Hayati Exact Paper In Compilation (EPIC) Jurnal Kedokteran Hewan Journal of Fisheries & Marine
Win Darmanto
Departement Of Biology, Faculty Of Science And Technology, Airlangga University, Indonesia
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Computer Science & IT Control & Systems Engineering Decision Sciences, Operations Research & Management Earth & Planetary Sciences Economics, Econometrics & Finance Education Engineering Environmental Science Health Professions Immunology & microbiology Industrial & Manufacturing Engineering Languange, Linguistic, Communication & Media Law, Crime, Criminology & Criminal Justice Materials Science & Nanotechnology Medicine & Pharmacology Nursing Public Health Social Sciences Veterinary Other

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Journal : Indonesian Journal of Biotechnology

 1 
Expression and purification of recombinant coat protein of sugarcane mosaic virus from Indonesian isolate as an antigen for antibody production Natalia Tri Astuti; Nurmalasari Darsono; Suvia Widyaningrum; Widhi Dyah Sawitri; Sri Puji Astuti; Win Darmanto
Indonesian Journal of Biotechnology Vol 24, No 1 (2019)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (72.263 KB) | DOI: 10.22146/ijbiotech.45551

Abstract

Sugarcane mosaic virus (SCMV, genus Potyvirus, family Potyviridae) is a prominent pathogen of sugarcane (Saccharum sp. hybrids). It can cause losses in susceptible varieties, in crop as well as sugar production, economically. Although it has been studied in major sugar-producing countries, research on the definement of SCMV from Indonesian isolates based on molecular study has been very limited. This study aimed to obtain a proper recombinant antigens emanating from coat protein of SCMV from Indonesian isolate in order to produce polyclonal antibodies that cann be used for immunodiagnosis assays in a subsequent study. A gene-encoding coat protein of SCMV (CP-SCMV) was amplified using RT-PCR and cloned into vector pJET1.2. The cDNA was inserted into 6X His-tag expression plasmid of pET28a(+) and over-expressed in Escherichia coli BL21(DE3) to produce a recombinant protein. The highest expression was found in 0.1M IPTG induction media for 5 h at 37oC. SDS-PAGE analysis clarified that the recombinant CP-SCMV remained as an insoluble fraction. Purifications was carried out by the affinity Ni-NTA resin, followed by electroelution to obtain a highly purified protein. To meet the quality requirements of a proper antigen, the highly purified protein was concentrated. A molecular weight of the rCP-SCMV (approximately 40 kDa) was clearly observed by 10% SDS-PAGE at the concentration of 16.184 mg/mL. 
The efficacy of a chicken antibody for the development of immunoassay‐based rapid detection in sugarcane mosaic virus disease Nurmalasari Darsono; Widhi Dyah Sawitri; Retnosari Apriasti; Agus Heri Setyo Wahyudi; Putri Andreyna Saragi; Victorin Mega Putri; Sugiharto Sugiharto; Win Darmanto
Indonesian Journal of Biotechnology Vol 28, No 1 (2023)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.74104

Abstract

Sugarcane Mosaic Virus (SCMV) infection is one of the most serious problems that can result in severe yield loss of sugarcane. Since the symptoms of SCMV infection are similar to other biotic and abiotic stress symptoms, the development of a rapid diagnostic with high precision is required. The use of laboratory animals such as rabbits is required for antibody production in immunoassay‐based detection. However, due to its many advantages, specific chicken egg yolk immunoglobulin (IgY) has received considerable attention as an alternative antibody production in immunodiagnostics for infectious diseases. In this study, IgY antibody against SCMV recombinant coat protein (CP) was successfully obtained from chicken blood serum and tested to compare its efficacy against antibody from rabbit (IgG) using immunocapture reverse transcription‐polymerase chain reaction (IC‐RT‐PCR). The result showed that IgY and IgG could detect 0.1 g SCMV infected leaves using 1000‐times‐diluted antibodies. The IgY antibody was also confirmed to be reproducible and potentially applicable in plant disease diagnostics using an antibody‐based detection.
Co-Authors A, Aulanniam A. Shofy Mubarak Agoes Soegianto Agus Abadi Agus Heri Setyo Wahyudi Alifah Hayati Andita Ayu Mandasari Aulanni'am, Aulanni'am Bambang Poernomo Betti Lukiati Betti Lukiati Binti Yunaida Djoko Agus P. Dwi Winarni Eko Prihiyantoro Eko Prihiyantoro Endah Rochmatika Endang Dewi Masithah farach Harianto Notopuro Herdiansyah, Mochammad Aqilah I Gede Widhiantara I.B. Rai Pidada Intan Permata Putri Intan Permata Putri Is Yuniar Juliana Christyaningsih Kusala, Muhammad Khaliim Jati Kusala Lailatul Lutfiyah, Lailatul Listijani Suhargo, Listijani Mammed Sagi Manikya Pramudya Mochammad Amin Alamsjah Mufasirin Mutiarawati, Diah Titik Nadyatul Ilma Indah Savira Nanik Siti Aminah Natalia Tri Astuti Nunuk Dyah Retno Lastuti Nurmalasari Darsono Nurmalasari Darsono P Kumaladewi Panjaitan, Novaria Sari Dewi Panjaitan Permatasari, Anak Agung Ayu Putri Pieter Kakisina Pujiastuti Wahyuningsih , Sri Puspita Sari, Evi Putri Andreyna Saragi Putu Angga Wiradana Ramadhana Ramadhana Retnosari Apriasti Ria Harmonis Rochmatika, Endah Rr. Juni Triastuti Samekto Wibowo Sapto Andriyono Sari, Ni Kadek Yunita Sari Siti Rahayu Sri Kadarsih Soedjono Sri Puji Astuti Sri Puji Astuti Wahyuningsih, Sri Puji Astuti Sugiharto Suhendra Agung Wibowo Sukardiman Susanto, Awaluddin Susilowati, Rina Priastini Suvia Widyaningrum Teguh Hari Sucipto, Teguh Hari Tri Dewanti W. Victorin Mega Putri Wiartini, Ni Wayan Ayu Wiartini Widhi Dyah Sawitri YULIA IRNIDAYANTI Yuni Kilawati