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Cloning and expression of NS2B/NS3 protein of DENV3 Indonesia strain in Saccharomyces cerevisiae expression system for the development of Dengue vaccine ASRI SULFIANTI; SABAR PAMBUDI; ISMA NUR AZIZAH; DODDY IRAWAN SETYO UTOMO; ABINAWANTO ABINAWANTO
Microbiology Indonesia Vol. 12 No. 2 (2018): June 2018
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (3936.725 KB) | DOI: 10.5454/mi.12.2.4

Abstract

NS3 protein is 618 amino acids (aa) in length containing serine protease and helicase domains required for DENV replication. Alignment of consensus amino acid sequences from all four DENV serotypes demonstrated that this protein is more conserved (78%) among the different dengue serotypes, which elicits a strong cellular immune response after viral infection in humans and animal models. Present study, a central hydrophilic region of NS3 cofactor, NS2B (NS2BH) with full length of NS3 genes DENV3 Indonesian strain were amplified from CDNA following PCR, and inserted to PYES2/CT shuttle vector. The recombinant plasmid was transformed and expressed in Saccharomyces cerevisiae expression system. As result, detection with Anti-His detector and Anti-NS3 shown NS2BH/NS3 was expressed as 83 KDA protein band. We found that addition of NS2BH on NS3 full length construction plasmid increase the yield and activity of protein expression in S. cerevisiae. In future study, our recombinant NS2B/NS3 protein can be used as recombinant protein in Dengue vaccine development.
Isolation of a Functional Gene Encoding Homologous Lysophospholipase from Indonesian Indigenous Bacillus halodurans CM1 SHANNI FERNANDA; ABINAWANTO ABINAWANTO; IS HELIANTI
Microbiology Indonesia Vol. 13 No. 1 (2019): March 2019
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (964.297 KB) | DOI: 10.5454/mi.13.1.2

Abstract

Lipase is a biocatalyst widely used in industry, for example detergent, pharmaceutical, food, or oil purification. One of the most widely lipase used for oil purification is lysophospholipase. As much as 50% of industrial enzyme needs are supplied from microorganisms. However, enzyme productivity from wild type microbial strain is usually limited and not applicable in industry, so that genetic engineering is necessary. Cloning gene encoding for lysophospholipase from Aspergillus niger and Cryptococcus neoformans have been conducted, but has never been conducted from alkalothermophilic bacteria, such as Bacillus halodurans. Bacillus halodurans CM1 is an alkalothermophilic bacterial strain isolated previously that has many industrially potential enzymes. This study aimed to isolate one of the gene encoding lipase from Bacillus halodurans CM1 and cloned into Escherichia coli DH5α using the pGEM-T easy vector. The gene fragment encoding lysophospholipase obtained with size 783 base pairs and had 100% similarity with gene encoding lysophospholipase from Bacillus halodurans C-125 (No access GenBank: BA000004.3). E. coli harbouring the recombinant plasmid with the gene also showed activity on trybutiryn medium compared to negative control.
The Effect of Cryopreservation on Cytochrome Oxidase1 (CO1) Gene and the Relationship with Spermatozoa Motility of Albino Pangasius catfish (Pangasionodon hypophthalmus) Uswatun Hasanah; Abinawanto; Alimuddin; Arief Boediono; Eni Kusrini
HAYATI Journal of Biosciences Vol. 30 No. 5 (2023): September 2023
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.30.5.825-833

Abstract

Cryopreservation is a technique for storing cells and tissues at very low temperatures for the possible usage of the stored cells and tissues throughout the year. Sperm cell cryopreservation in some species causes a decrease in sperm quality and DNA damage. Inappropriate cryopreservation protocols can cause changes in sperm physiology. Mitochondria are organelles that play a role in producing energy for sperm motility. Mitochondria have DNA molecules with small sizes compared to structures from nuclear DNA. This study analyzed the effect of cryopreservation on sperm motility and the Cytochrome Oxidase1 (COI) gene. The CO1 gene in mitochondrial DNA plays a role in energy production for spermatozoa motility. The cryopreservation was performed using skim milk and 10% methanol cryoprotectant, and the temperature in the equilibration process was 4-5°C for 10 minutes. Cryopreservation took place for 14 days in the freezer at -80°C. In addition, the thawing process was performed for 1-2 minutes at 40°C. This study found that the number of lesions per 10 kb in the CO1 gene in post-equilibration spermatozoa was (9.24±3.74), and post-thawing spermatozoa (was 10.26±7.54). Spermatozoa motility was obtained in fresh spermatozoa, i.e., 87±1.5%, post-equilibration spermatozoa 79±4.5%, and post-thawing spermatozoa 30±3.2%. This study concluded that cryopreservation of spermatozoa causes CO1 gene lesions and that in cryopreserved spermatozoa, there is a decrease in spermatozoa motility compared to fresh spermatozoa.
Detection of Submicroscopic Soil-Transmitted Helminth Infections from Fecal Samples in Nangapanda, Ende, Using Real-Time Polymerase Chain Reaction Maylasari, Roospita; Wulandhary, Sendy; Supali, Taniawati; Abinawanto, Abinawanto
Makara Journal of Science Vol. 18, No. 2
Publisher : UI Scholars Hub

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Abstract

Soil-transmitted helminth (STH) infections (Necator americanus (hookworm), Ancylostoma duodenale (hookworm), and Ascaris lumbricoides) can lead to anemia, malnutrition, and iron deficiency. Traditionally, STH infections have been diagnosed using microscopy to detect eggs in human fecal samples. However, there are several limitations of this method. The aim of this research was to detect the percentage of submicroscopic STH infections from human fecal samples (children, 5–18 years old) in Nangapanda, Ende, using the real-time polymerase chain reaction (PCR) method. The fecal samples were collected in two time periods, which were before and after treatment, using 400 mg of Albendazole. There were 242 samples in total, but only 45 negative samples from microscopic detection were tested with real-time PCR. The DNA samples were isolated and amplified wih primers of internal transcribed spacer (ITS-1 and ITS-2) region of rDNA. The detection of samples with real-time PCR generated an amplification curve in VIC, FAM, and Texas Red fluorophore. Three samples (6.7%) in pre-treatment were low load of DNA (N. americanus and A. lumbricoides) (Ct > 35). Four samples (9.1%) were low load of DNA (N. americanus) (Ct > 35) in post-treatment. Five samples (11.4%) were moderate load of DNA (A. lumbricoides) (30 < Ct < 35) in post-treatment. This study showed that real-time PCR could detect submicroscopic infections from specific species of hookworm and A. lumbricoides
Detection of Papua New Guinea Thalassemia Alpha Mutation in Gayo, Sumba, Ternate, and Timika Populations Nurfitriani, Risya; Abinawanto, Abinawanto; Noviyanti, Rintis; Trianti, Lely; Nainggolan, Ita M.
Makara Journal of Science Vol. 18, No. 2
Publisher : UI Scholars Hub

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Abstract

Papua New Guinea (PNG) mutation is a point mutation that occurs in noncoding region of alpha globin clusters. Polymorphism promotes an additional recognition site for transcription factor (GATA-1) which presumably downregulates alpha globin synthesis. The aim of this research is to detect PNG mutation in other populations in Indonesia, thus the results will be used for completing standard diagnoses in detecting alpha thalassemia mutation based on ethnic background. The method used in detecting PNG mutation was PCR-RFLP. Detection of 399 samples (MCH
Effect of Oral Administration of Dehydroepiandrosterone on PCOS-Like Phenotype of Female C57BL/6 Mice Sundari, Ayu Mulia; Prakoso, Nurul Muhammad; Dwiranti, Astari; Abinawanto, Abinawanto; Boediono, Arief; Funahashi, Hiroaki; Bowolaksono, Anom
Jurnal Sain Peternakan Indonesia Vol 19 No 2 (2024)
Publisher : Universitas Bengkulu

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.31186/jspi.id.19.2.111-116

Abstract

We aim to evaluate the efficacy and optimal dose of orally administered DHEA in the PCOS mice model by assessing their ovarian morphology and serum FSH, LH, and testosterone levels. Female C57BL/6 mice were divided into a control group (n=5, received daily injections of 0.2 ml sesame oil) and an experimental group, which was further classified into 1) D-45 group (n=5), 2) D-60 (n=5), and 3) D-90 group (n=5) that were treated with 45, 60, and 90 mg/Kg body weight of oral DHEA. After modelling, mice in the control group had a regular estrous cycle, while mice in all experimental groups had a disturbed estrous cycle. Ovarian histology showed several growing follicles and some corpora lutea (CL) in the control, D-60, and D-90 groups. However, some large antral follicles and decreased CL were observed in the D-45 group. Serum FSH was significantly lower in the D-45 group than in the control group (3.73 ± 0.12 vs. 5.28 ± 0.31 mIU/mL, P<0.01), but D-60 and D-90 groups had a similar FSH level to the control group (P>0.05). The serum level of LH and testosterone were significantly elevated in the D-45 group than in the control group (2.52 ± 0.43 vs. 1.30 ± 0.33 mIU/mL, P<0.01 and 1.80 ± 0.32 vs. 1.24 ± 0.23 ng/mL, P<0.01, respectively). Still, the level of LH and testosterone in the D-60 and the D-90 groups was comparable to the control group (P>0.05). Our study suggests that oral administration of DHEA is efficacious in establishing PCOS-like phenotype in mice with the suggested optimal dosage of 45 mg/Kg body weight.
Optimizing the Expression of Polyethylene Terephtalate Hydrolase-Encoding Synthetic Gene in Escherichia coli Arctic Express (DE3) Nataniel, Jocelyn; Ulfah, Maria; Achnafani, Dini; Nurhayati, Niknik; Sabbathini, Gabriela Christy; Wulandari, Sri Rezeki; Abinawanto, Abinawanto; Helianti, Is
Makara Journal of Science Vol. 28, No. 2
Publisher : UI Scholars Hub

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Abstract

The waste of polyethylene terephthalate (PET) plastic waste in Indonesia is a pressing concern due to its slow degradation and potential environmental damage. One promising solution is to utilize polyethylene terephthalate hydrolase from Ideonella sakaiensis (IsPETase), an enzyme that specifically degrades PET. However, inducing the expression of IsPETase synthetic gene in Escherichia coli BL21 (DE3) has been challenging because much of it remains insoluble. This study aimed to express IsPETase in E. coli Arctic Express (DE3) and optimize the conditions to enhance its production. First, pET22b(+)pelB-IsPETase was inserted into E. coli Arctic Express (DE3). The recombinant E. coli Arctic Express (DE3) was induced with isopropyl-β-D-1-thiogalactopyranoside (IPTG) and incubated at 10 °C. The fraction expressing soluble IsPETase was determined in different culture media, IPTG concentrations, induction times, and soni-cation durations. Parameters were optimized using a one-factor-at-a-time approach and then evaluated based on esterase specific activity and SDS-PAGE analysis. Results showed that IsPETase can be expressed in extracellular, periplasmic, and cytoplasmic soluble fractions. However, the extracellular fraction should be concentrated. Subsequent optimization focused only on the cytoplasmic fraction under optimal conditions, achieving a threefold increase in PETase specific activity compared with that under uninduced IPTG conditions. The reaction of PETase enzyme with PET and PCL was proven by weight loss, Scanning electron microscope (SEM), and Fourier transform infrared spectroscopy (FTIR). Although successful IsPETase expression and production optimization have been achieved, the specific activity remains low, prompting the need for ongoing expression optimization.
Early Development Of The Tetra Cardinalfish: Paracheirodon Axelrodi Schultz 1956 Cindelaras, Sawung; Abinawanto, Abinawanto; Kusrini, Eni
International Journal of Islamic Education, Research and Multiculturalism (IJIERM) Vol 7 No 1 (2025)
Publisher : The Islamic Education and Multiculturalism Foundation

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.47006/ijierm.v7i1.417

Abstract

Cardinal tetra, Paracheirodon axelrodi, merupakan ikan hias yang populer di Indonesia dan mancanegara. Penyumbang ikan hias akuarium di Rio Negro, Amerika Selatan. Komoditas ekspor ikan hias ke Amerika, Jepang dan Singapura. Ikan ini sangat sulit dikembangbiakkan di lingkungan aslinya. Informasi ilmiah tentang reproduksi, perkembangan embrio dan larva masih sangat sedikit. Tujuan penelitian adalah untuk memperoleh informasi tentang reproduksi, perkembangan embrio dan larva cardinal tetra. Informasi ini sangat penting untuk meningkatkan perkembangbiakan ikan cardinal tetra dan konservasi ex situ. Sebanyak 20-50 butir telur digunakan untuk mengamati embrio melalui penetasan dengan mikroskop stereo pada perbesaran 50-100x. Embrio diamati dengan mikroskop stereo sebanyak 20-50 butir telur pada perbesaran 1-2,5x. Fekunditas berkisar antara 115 sampai 550 butir telur. Telur berbentuk bulat, transparan, agak lengket dan tenggelam ke dasar. Diameter telur 0,835-2,202 ±3,7 mm saat menetas dan terhidrasi. Panjang larva saat menetas 2,406-2,703 ±86,96 mm. Kuning telur habis setelah 4-5 hari. Bukaan mulut 0,235-0,366 mm cukup untuk memakan nauplii Artemia sp. Larva mulai berenang bebas setelah 4 hari. Perkembangan larva hingga tahap akhir memakan waktu 28-30 hari.Kata Kunci: Embrio, Larva, Perkembangan, Pertumbuhan, Pembiakan, Konservasi The cardinal tetra, Paracheirodon axelrodi, is a popular ornamental fish in Indonesia and abroad. Contributor to ornamental aquarium fish in Rio Negro, South America. Export commodity of ornamental fish to America, Japan and Singapore. This fish is very difficult for breeding in its native environment. There is little scientific information on reproduction, embryonic and larval development. The objective of the research was to obtain information on the reproduction, embryonic and larval development of the cardinal tetra. This information is essential to improve the breeding of cardinal tetra fish and ex situ conservation. A total of 20-50 eggs were used to observe embryos through hatching with a stereo microscope at 50-100x magnification. Embryos were observed with a stereo microscope for 20-50 eggs at 1-2.5x magnification. Fecundity ranged from 115 to 550 eggs. Eggs are round, transparent, slightly sticky and sink to the bottom. Egg diameter 0.835-2.202 ±3.7 mm at hatching and hydration. Larval length at hatching 2.406-2.703 ±86.96 mm. Yolk depleted after 4-5 days. Mouth opening of 0.235-0.366mm is sufficient for consuming the Artemia sp. nauplii. Larvae start swimming freely after 4 days. Larval development to the final stage takes 28-30 days.Keywords: Embryo, larva, development, growth, breeding, conservation
Co-Authors A. Sofyan A. Sofyan Abdul Basith Achmad Dinoto Achmad Dinoto Achnafani, Dini Ade Sunarma Agung Sudaryono Agus Haryadi Ahmad Aulia Jusuf Alimuddin Andi Salamah ANOM BOWOLAKSONO Arief Boediono ASRI SULFIANTI Bayu , Mariana D. Diki Nurul Huda Djamhuriyah S. Said DODDY IRAWAN SETYO UTOMO Dwiranti, Astari Endang C. Purba Endang Sukara Eni Kusrini Eni Kusrini Eni Kusrini Eni Kusrini, Eni Erma Primanita Hayuningtyas Fitrianingrum, Nisa Funahashi, Hiroaki H. Julendra Hidayat Hidayat Huriyah Adani Saoemi IS HELIANTI IS HELIANTI is helianti Isma Nur Azzizah Ita M. Nainggolan L. Istiqomah Lina Mulyawati Lulut Dwi Sulistyaningsih M. F. Karimy M. F. Karimy Maria Ulfah Maria Ulfah Mariana D. Bayu Marina Silalahi Marlina Ardiyani Maylasari, Roospita MILANI ANGGIANI Nataniel, Jocelyn NIKNIK NURHAYATI NIKNIK NURHAYATI Nisyawati Nova Anita Noviyanti, Rintis Nurfitriani, Risya Odang Carman Prakoso, Nurul Muhammad PUSPITA LISDIYANTI Putra Mahanaim Tampubolon Rabbil Pratama Aji Retno Lestari Retno Lestari Retno Lestari Retno Lestari Riska S. Wahyuningtyas Rita Rostika RONI RIDWAN Sabar Pambudi Sabbathini, Gabriela Christy Sanya Khaerunnisa Sawung Cindelaras, Sawung SHANNI FERNANDA Shanti Ratnakomala Siti Z Musthofa Solly Aryza Sulistiani ., Sulistiani Sulistiani Sulistiani Sulistiani Sulistiani, Sulistiani Sunarma , Ade Sundari, Ayu Mulia Taniawati Supali Trianti, Lely Uswatun Hasanah UTAMA ANDI Valerry Athalia Priyanka Wibowo Mangunwardoyo WIBOWO MANGUNWARDOYO Wulandari, Sri Rezeki Wulandhary, Sendy Yasman Yasman Yasman Yushinta Fujaya