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All Journal HAYATI Journal of Biosciences Jurnal Ilmu dan Teknologi Kelautan Tropis ESENSI: JURNAL BISNIS DAN MANAJEMEN ETIKONOMI Islamadina : Jurnal Pemikiran Islam ILMU KELAUTAN: Indonesian Journal of Marine Sciences Journal of Marine Research JURNAL ILMIAH MANAJEMEN & BISNIS TEKMAPRO Journal of Industrial Engineering and Management EQUILIBRIUM JMM17: Jurnal Ilmu Ekonomi dan Manajemen Indonesian Journal of Cancer Chemoprevention Media Trend: Berkala Kajian Ekonomi dan Studi Pembangunan Journal of Consumer Science UNEJ e-Proceeding Jurnal Ilmiah Ekonomi Islam Jurnal Manajemen International Journal of Artificial Intelligence Research Journal of Consumer Sciences Jurnal Ekonomi dan Bisnis EKONOMIS : Journal of Economics and Business J-MKLI (Jurnal Manajemen dan Kearifan Lokal Indonesia) JMKSP (Jurnal Manajemen, Kepemimpinan, dan Supervisi Pendidikan) JURNAL PENDIDIKAN TAMBUSAI International Journal of Supply Chain Management Ekono Insentif KOMIK (Konferensi Nasional Teknologi Informasi dan Komputer) Ekuilibrium : Jurnal Ilmiah Bidang Ilmu Ekonomi JETL (Journal Of Education, Teaching and Learning) ISOQUANT : Jurnal Ekonomi, Manajemen dan Akuntansi Jurnal Maksipreneur: Manajemen, Koperasi, dan Entrepreneurship JPM PAMBUDI Almana : Jurnal Manajemen dan Bisnis Jurnal Akuntansi dan Bisnis (JRAMB) Jurnal Riset Akuntansi Mercu Buana CAM JOURNAL: Change Agent For Management Journal Jemasi Jurnal Ekonomi Manajemen dan Akuntansi Jurnal Manajerial Jurnal Ilmiah Edunomika (JIE) Jurnal Riset Entrepreneurship ASSET: Jurnal Manajemen dan Bisnis Masyarakat Berdaya dan Inovasi Budimas : Jurnal Pengabdian Masyarakat Abdimasku : Jurnal Pengabdian Masyarakat ANALISIS - SCIENTIFIC JOURNAL OF ECONOMICS, MANAGEMENT, BUSINESS, AND ACCOUNTING UTAMAX : Journal of Ultimate Research and Trends in Education AKTUAL Community Empowerment Jurnal Manajemen dan Inovasi (MANOVA) Journal of Entrepreneurship & Business Jurnal Valuasi : Jurnal Ilmiah Ilmu Manajemen dan Kewirausahaan Jurnal Bisnis dan Manajemen (JBM) Jurnal Riset dan Aplikasi Mahasiswa Informatika (JRAMI) Society: Jurnal Pengabdian Masyarakat Bussman Journal : Indonesian Journal of Business and Management Jurnal Ekonomi, Manajemen, Bisnis dan Akuntansi (JEMBA) BISMA (Bisnis dan Manajemen) Makara Journal of Science Makara Journal of Technology Innovative: Journal Of Social Science Research Sosiologia : Jurnal Agama dan Masyarakat Indo-Fintech Intellectuals: Journal of Economics and Business Jurnal Pendidikan, Kebudayaan dan Keislaman JIM: Jurnal Ilmiah Mahasiswa Pendidikan Sejarah JPM Pambudi IQTISHADUNA: Jurnal Ilmiah Ekonomi Kita Indonesian Journal of Forestry Research Indonesian Journal of Jamu J-CEKI Jurnal Ekonomi, Manajemen, Akuntansi OPTIMA Jurnal Penelitian Hasil Hutan ILJS Annales Bogorienses
Adi Santoso
Universitas Telkom
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Journal : Annales Bogorienses

 1 
An Application of Reverse Transcriptase Polymerase Chain Reaction in A Relative Quantification of Gene Expression Santoso, Adi
Annales Bogorienses Vol. 9 No. 1 (2004): Annales Bogorienses
Publisher : BRIN

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The ability to quantify steady state levels of individual messenger-RNA (mRNA) transcripts has been the key issue for study on the control of gene expression. Although two available techniques, Northern blot and nuclease protection assays (NPA) have been widely used tor detecting mRNA, these techniques have critical limitations. The most obvious limitation of these two techniques is the required number of target mRNAs to be detected. Reverse transcription-polymerase chain reaction (RT-PCR), which has been accepted as a highly sensitive and specific method, provides a means for detecting and quantifying gene expression using, theoretically only a single molecule of mRNA. The sensitivity and reliability of RT-PCR is dependent upon both the RT and PCR steps. The PCR step has been problematic because of the exponential nature of this reaction where small variation can lead to dramatic changes in final result. Therefore, the use of RT-PCR for quantification of gene expression requires pre-experimental planning and design. In this experiment, the procedure for pre-experimental planning, linear range determination and subsequent relative quantification of gene expression are described in detail. A study of ornithine decarboxyJase gene, a gene involved in the polyamine biosynthesis and temporally expressed, during embryogenesis of Musca domestica (housefly) was used as the model. The results show that during early embryogenesis (t-1 to t-4) the expression level was very low. The increase in expression profile was observed started at t-5, peaked at t-9, and followed by substantial decrease from t-10 to t-12.
Optimization Expression and Stability Test of Recombinant Human Interferon Alfa 2a Fusion Protein in Escherichia coli BL21 (DE3) Santoso, Adi; Kusumawati, Arizah
Annales Bogorienses Vol. 19 No. 1 (2015): Annales Bogorienses
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The rhIFN α2a is expressed as a fusion protein containing thioredoxine and polyhistidine sites at its N terminal. Our previous research has obtained recombinant human IFN α2a (rhIFN α2a) protein that expressed predominantly as a soluble form in E. coli BL21 (DE3). Through systematic approach of various culture conditions, the aim of current this research is to acquire the best condition and its stability of recombinant rhIFN α2a fusion protein in a culture under study. Expression optimization performed by using three parameters, i.e.: temperature, induction time and inducer concentration. Various IPTG concentrations are 0.25, 0.5, 0.75, and 1.0 mM. The incubation time of bacterial cell culture carried out in 3, 4, and 5 hours at temperature 28, 30, and 37°C. The best condition was used to analyze the stability of rhIFN α2a protein expression up to ten generation. The expressed protein was analyzed using SDS PAGE and CBB staining. The optimal culture condition was found to be 37 °C temperature with 4 hours time of induction and 1 mM IPTG concentration. Stability analysis revealed that the rhFN α2a protein expression remained stable until the tenth generation with molecular weight, approximately, 36 kDa. 
Optimizaton of Cationic Lipid Mediated Transfection of pEGFP-c1 and pJ-EPO Plasmids in Chinese Hamster Ovary (CHO) Cells Attached Culture for Transient and Stable Recombinant Human Erythropoietin (rhEPO) Expression Septisetyani, Endah Puji; Kusumawati, Arizah; Santoso, Adi
Annales Bogorienses Vol. 19 No. 1 (2015): Annales Bogorienses
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Cationic lipid is one of transfection agents which show high efficiency and low cytotoxicity. The transfection efficiencies can be varied depending upon the type or amount of cationic lipids, the cell line or DNA plasmid being used for transfection. The purpose of this study was to find optimal condition for transfection of CHO-K1 and CHO-S cells with pJ-EPO plasmid (containing human erythropoietin/ hEPO gene) compared with pEGFP-c1 plasmid (containing green fluorescence protein/gfp gene) by cationic lipid Lipofectamin 2000TM (lipofectamin) to generate stable transfectant expressing recombinant human erythropoietin (rhEPO). Optimization was carried out regarding the amount of lipofectamin, DNA concentration, and concentration of antibiotic Geneticin (G418) for selection of stable transfectants. By using standard amount of lipofectamin (10 μl/well) in 6-well plate, highest expression level of green fluorescent protein (GFP) was shown after transfection of CHO-K1 cells with 3 μg/well pEGFP-c1 while highest expression level of rhEPO was observed after transfection of CHO-K1 cells with 6, 8, or 10 μg/well pJ-EPO plasmid. The data also indicated that optimal transfection conditions of CHOK1 and CHO-S cells with pJ-EPO were shown with the use of 4 μg/well DNA in combination with 15 μl lipofectamin. Concentration of G418 used during cells selection also affected the expression where strongest rhEPO expression was shown at 750 ng/μl G418 concentration. Similar to GFP expression profile, rhEPO signal was detected very low during selection process sbased on Western blot data at day 9. Stronger rhEPO signal was observed after day 20 when the stable transfectants have been obtained.
In-Silico Cloning and Analysis of Divalent Subunit OMP31-SODc Proteins As A Prophylaxis Vaccine Against Brucella melitensis Infection Wijaya, Sri Kartika; Kusumawati, Arizah; Wardiana, Andri; Rubiyana, Yana; Husnaa, Ulfatul; Santoso, Adi
Annales Bogorienses Vol. 19 No. 1 (2015): Annales Bogorienses
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The urgency to develop a new protein subunit based vaccine candidate against Brucella was provoked by its frequent infection to human and livestock. Since Brucella melitensis is found as the most frequently isolated Brucella species from human, thus the outer membrane of B. melitensis becomes a prominent subcellular localization to search for promising antigen to be developed as vaccine candidate due to its interaction with host cell. Among outer membrane proteins suggested by Vaxign program, OMP31 was found as the most promising candidate. Moreover, analysis on other subcellular localization led our interest to SODc protein, which was expected to support OMP31 in triggering immune response. The OMP31-SODc divalent vaccine candidate was analysed in silico to predict its stable three-dimensional structure, cloning process and expectation on the ease during expression, purification and vaccine delivery to elicit the expected immune response.
Sequential Adaptation in Mammalian CHO-K1 Cells Producing Human Erythropoietin Wisnuwardani, Popi Hadi; Septisetyani, Endah Puji; Santoso, Adi
Annales Bogorienses Vol. 21 No. 1 (2017): Annales Bogorienses
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Abstract

The production of recombinant proteins for clinical applications using mammalian cell technology has become a prevalent system because of its capacity in assembling functional proteins. One of the main problems with CHO-K1 cells is that this cell has to grow in the presence of serum. However, the presence of serum will complicate the downstream step for protein production. Thus, protein produced in media without serum, theoretically, would be easier to purify. Technically, this type of cell can be produced by growing the CHO-K1 cells in serum-free media by using adaptation method in suspension condition. This research showed that through sequential adaptation using conditioned media, the CHO-K1 cell line that produces the human erythropoietin gene (hEPO) was able to grow in suspension culture using serum-free media. Based on Western blot analysis, it showed that the protein (hEPO) was able to be expressed in suspension culture with molecular mass of about 47 kDa.
Heterologous Expression of Recombinant Human Insulin Glargine (hIG) in Methylotrophic Yeast Pichia pastoris Herawati, Neng; Rubiyana, Yana; Desriani, Desriani; Santoso, Adi
Annales Bogorienses Vol. 26 No. 1 (2022): Annales Bogorienses
Publisher : BRIN

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ann.bogor.2022.v26.n1.13-20

Abstract

World health organization (WHO) announced that diabetic patients increased significantly yearly worldwide. Consequently, the need for insulin becomes very large. Pichia pastoris, defined as methylotrophic yeast and a well-known expression and protein production host, is widely used for biopharmaceutical-based drug production. This research aims to synthesize human insulin glargine (hIG) protein in yeast P. pastoris. Human insulin glargine is a group of long-acting analogue insulin.We used a synthetic hIG-encoding gene constructed in frame with the truncated α-factor secretion signal in a pD902 expression vector. Recombinant plasmid pD902-hIG was linearized using Sac1 enzyme and transformed into P. pastoris genome. Multicopy clones were selected on YPDS plates containing Zeocin™ with concentrations of 100; 500; 1,000; and 2,000 µg//mL. Analyses using SDS-PAGE, slot blot, and Western blot showed that recombinant hIG protein had been obtained with a molecular weight of approximately 6,000 Daltons.
A Preliminary Report on The Syntheses of Oligonucleotide Primers in The National Research and Innovation Agency (NRIA) Atikana, Akhirta; Prasetyoputri, Anggia; Rubiyana, Yana; Herawati, Neng; Desriani, Desriani; Pratiwi, Riyona Desvy; Wulandari, Dwi; Sukmarini, Linda; Kusharyoto, Wien; Santoso, Adi; Putra, Masteria Yunovilsa; Lisdiyanti, Puspita
Annales Bogorienses Vol. 26 No. 1 (2022): Annales Bogorienses
Publisher : BRIN

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ann.bogor.2022.v26.n1.21-27

Abstract

A PolyGen DNA Synthesizer is equipment that is used for synthesizing oligonucleotide primers for any amplification targets. Oligonucleotide primers are indispensable components for any Polymerase Chain Reaction (PCR)-based detections. In the present study a number of oligonucleotide primer sets were synthesized to target (1) the Human Insuline Glargin (HIG) and (2) the Human Erythropoietin (EPO), as well as (3) the RNA-dependent RNA Polymerase (RdRp) and (4) the Nucleocapsid (N) genes of the severe acute respiratory syndrome virus 2 (SARS-CoV-2). A solid-phase oligonucleotide synthesis method was used according to the default protocol of the Polygen’s instrument to synthesize primers at a 40 nmol scale. The synthesized primers in this study were compared to commercially produced primers in their ability to amplify the gene target(s) in PCR and quantitative real-time PCR (qPCR) reactions. The first two sets of primers showed similar results in PCR compared to commercial primers; however, these primers were not tested for qPCR due to sample limitation. In contrast, the primer sets 3 and 4 were not able to produce amplicons in PCR reactions and only the primer set 4 successfully amplified the gene target in qPCR. These results indicate that the crude primers synthesized in this study are promising candidates for molecular detection and diagnostics, but these primers would benefit from further optimization for routine applications.
Co-Authors Abdurohim, Dindin Aghnia Salma Salsabila Alberta Adinata Alghazeer, Rabia Alifah Febirika Nurjanah Amelia Putri Amellya, Risnanda Putri Anam, M. Khairul Ananda, Yudha Rizky Andri Wardiana, Andri Anggorowati, Luki Sri Anggraeni, Fahrida Noor Anugerah Yuka Asmara Aprylia, Regyta Ardianto*, Felix Ardyan Firdausi Mustoffa Ari Irawan Astuti, Nanda Widia Atikana, Akhirta Aulia, Amanda Karismatul Auliasari, Ria Windi Azhar, Nuril Azzahra, Salsabila Bafirman HB Bambang Yulianto Beltasari, Ruri Acnes Bety Oktavia Rahayu Chamidah, Siti Chamidah, Siti Chandra Bella Kartika Chrisna Adhi Suryono Dede Hermawan Delianis Pringgenies Desriani Desriani Dwi Astuti Dwi Warni Wahyu Dwi Wulandari Dwisakti*, Vebrian Edi Kurniawan Edy Kurniawan, Edy Efendi, Zakaria Eka Destriyanto Pristi Eka Destriyanto Pristi Eka Destriyanto Pristi Ayuningtyas Endah Puji Septisetyani, Endah Puji Ervia Yudiati Evit Harmawati Fatimah Azzahra Febrianti Muji Trianawati Felix Ardianto* Fery Setiawan Fery Setiawan Fery Setiawan Fery Setiawan Fery Setyawan Fery Setyawan Fitri Suratin Fitria Hidayati, Fitria Gessy Augustina Dwi Saputri Hadi Sumarsono Hadi Sumarsono Hanik Rahmawati, Hanik Hanim, Meysella Al Firdha Happy Fitria, Happy Heri Wijayanto Herma Rio Candra Saputra Himawan, Alayna Lillahida Indri Husnaa, Ulfatul Iin Wijayanti Indrawan , Ahmad Faras Iqomatuddin, Muhammad Irmanida Batubara Isna Nur Rahmawati Ita Widowati Juliawati Juliawati Kartika, Chandra Bella Kastian, Ria Fajarwati Khasanah, Alfina Asma’ul Kiswanti, Nafisyah Komang Alit Paramitasari Kristyana, Naning Kumala, Masria Kusharyoto, Wien Kuska, Dila Ayu Ramanda Kusumaningrum, Reny Kusumawardhani, Dhian Kusumawati, Arizah La Ode Sugianto Laila Fijannatin Naim Laksono Trisnantoro Lestari, Marlina Dewi Lina Pina Uljannah LINDA SUKMARINI M. Sulastiningsih, Ignasia Muhammad Rizky Alamshah Mulyadi Mulyadi Naning Kristiyana Naning Kristiyana Naning Kristiyana Naning Kristiyana Naning Kristiyana, Naning Nayni Nayni Neng Herawati, Neng Nisa'u Lailatizzahro Nistirohah Nur Fitriani Niwayansari, Fira Norma Hapsary Novianti, Metta Nuraini , Ria Azizah Tri Nurdiana Nurdiana Nurul Iman Nurul Iman, Nurul P Cahyono Pari, Rohmah Popi Hadi Wisnuwardhani, Popi Hadi Pradana, Sepbrio Ian Pradipta, Farel Bragas Pramudya, Dimas Alfikri Prasetyaningrum, Pekik Wiji Prasetyo, Ilham Adi Prasetyoputri, Anggia Pratama, Dimas Dipa Pratiwi, Anggun Anggraini Mila Pristi, Eka Destriyanto Purnomo, Rochmat Aldy Purwaningrum, Titis Purwantiningsih Sugita PUSPITA LISDIYANTI Puspita Sari Puspitasari, Muthia Fahriyah Putra, Masteria Yunovilsa Putri, Isna Hary Ardita Radita Noviani Rahayu, Listyowati Puji Ramadani, Ratna Dwi RATIH ASMANA NINGRUM Ratri Widya Wulandari REIGION JUMANTORO Renaldi, Bagas Revina Lindya Kusuma Riawan Riawan Riawan Riawan Riawan Riawan, Riawan Riski Taufik Hidayah Ristianawati, Yuyun Rivaldi, Hengky Riyona Desvy Pratiwi, Riyona Desvy Riza Dessy Nila Ayutika Rizki Gagah Saputra Rudhi Pribadi Rusdjijati, Retno Ryza Wahyu Anofa S Hartono Sabela, Rizka Elis Safitri, Unna Ria Salsabila, Puti Athirah Sari, Lina Fatika Sepbrio Ian Pradana Setiawan Edi Wibowo Setya Raharja Shabrina, Yuniar Nur Siska Maryunitasari Siti Arifah Sri Hartono Sri Hartono Sri Hartono, Sri Subarkah, Windu Praditiyo Suhaimi, Ahmad Sunarya, Yaya Sunaryo Sunaryo Susanto, Akhiyan Hadi Suyoko, Ahmad Tasya Melania Nasution Titi Rapini Titi Rapini, Titi Titis Purwaningrum Tsabita, Azza Salma Uljannah, Lina Pina Umi Farida Umi Farida Umi Farida Umi Farida Vebrian Dwisakti* Vicamara, Ulfa Wahna Widhianingrum Wahna Widhianingrum Wahna Widhianingrum, Wahna Wahyuningsih, Dwi Warni Wayan Wijaksono Wibowo, Rahmada Yasmin Ari Wijaksono, Wayan Wijaya, Sri Kartika Wijianto Wijianto Wisnasari, Shila Wisnuwardani, Popi Hadi Yana Rubiana, Yana Yana Rubiyana, Yana Yulian Wiji Utami Yuliana Mariya Ulva Yuliarta, Refina Dwi Zeini, Cerlis Cesar