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All Journal Microbiology Indonesia Annales Bogorienses
ASRI SULFIANTI
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Biochemistry, Genetics & Molecular Biology Immunology & microbiology Medicine & Pharmacology

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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.
The Potency of Aluminum Hydroxide Nanoparticles for Dengue Subunit Vaccine Adjuvant SABAR PAMBUDI; ETIK MARDLIYATI; SILMI RAHMANI; DAMAI RIA SETYAWATI; TIKA WIDAYANTI; ANGELINA GILL; ASRI SULFIANTI; WHINIE LESTARI
Microbiology Indonesia Vol. 12 No. 3 (2018): September 2018
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (984.8 KB) | DOI: 10.5454/mi.12.3.5

Abstract

The potency of aluminum hydroxide as an adjuvant in vaccine development is considered to depend on its particle size. In previous studies, we have successfully prepared two size particle, micro, and nano, aluminum hydroxide gel (alum) adjuvants. The potency of those particles as a candidate of adjuvant is needed to be characterized. In this study, we formulated our adjuvants with purified DENV3 pre Membrane Envelope (prM-E) recombinant protein and evaluated the induction of nitric oxide level in mouse macrophage RAW 264.7 cells. We prepared the alum adjuvant by precipitation-homogenization methods with an agitation rate at 11,000xg. Secreted prM-E recombinant protein was collected from Pichia pastoris X-33 fermentation which produced using bioreactor. Recombinant protein purification was carried out by anion exchange chromatography followed with size exclusion chromatography. The purified prM-E recombinant protein was observed as a single band around 70 -1k Da with a concentration of 105 mg mL . Complex nanoparticles alum with prM-E protein significantly (p<0.05) induced the nitric oxide level. Further analysis should be conducted in order to discover the detail molecular mechanism of nanoparticle alum adjuvant, recombinant protein, and cellular immune response.
ANTI-DIABETIC EFFECT OF 70% ETHANOL EXTRACT OF Anredera cordifolia (Teen) Stennis (BINAHONG) LEAVES ON GLUCOSE TRANSPORTER 4 (GLUT4) AND PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR GAMMA (PPAR-γ) GENES EXPRESSION IN 3T3-L1 ADIPOCYTE CELLS Asri Sulfianti
Annales Bogorienses Vol. 25 No. 1 (2021)
Publisher : BRIN

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Abstract

The evidence of Anredera cordifolia (Binahong) as anti-diabetic at molecular stage has not been elucidated. Present study, we investigate the antidiabetic potential of Binahong by evaluating its effect on two diabetic gene markers expression, Translocation of Glucose Transporter 4 (GLUT4) and Peroxisome Proliferator-Activated Receptor Gamma (PPAR-γ) in 3T3-L1 adipocytes. The investigation has been done by semi quantitative real time PCR. Our finding shown, the GLUT4 MRNA levels were dose dependent excess in adipose treated by 6.25-12.5 mg/ml of 70% Binahong ethanol extract. Though, at 50 mg/ml dose, the GLUT4 expression was up down to 1.2 fold. In addition, the pattern of PPAR-γ MRNA levels in 3T3-L1 were almost same. Every Binahong extracts gave increment on PPAR-γ MRNA levels except 3.125, 6.25, and 50 mg/ml doses. In undifferentiated adipocytes, GLUT4 and PPAR-γ mRNA were barely expressed. The up regulation of those MRNA levels probably occurred because of Binahong has an active compound. However, it is not yet known which compounds who regulate the expression and induces translocation of GLUT4. We hypothesis that it acts as ligand for the PPAR-γ transcriptional factor which activates the GLUT4 transcription and plays orchestra in glucose homeostasis at molecular level.
Co-Authors Abinawanto Abinawanto ANGELINA GILL DAMAI RIA SETYAWATI DODDY IRAWAN SETYO UTOMO Etik Mardliyati Isma Nur Azzizah Sabar Pambudi SILMI RAHMANI TIKA WIDAYANTI Whinie Lestari