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Cloning and Expression of Nonstructural Protein NS1 of Dengue Virus Serotype 2 BETI ERNAWATI DEWI; FITHRIYAH FITHRIYAH; ANDRIANSJAH RUKMANA; PAISAL PAISAL; DEKA LARASATI; TJAHJANI MIRAWATI SUDIRO
Microbiology Indonesia Vol. 6 No. 1 (2012): March 2012
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (614.911 KB) | DOI: 10.5454/mi.6.1.3

Abstract

Early diagnosis of dengue virus (DENV) infection is affirmative for patient management and control of the disease. Detection of nonstructural-1 (NS1) antigen has been proven to provide early detection of DENV infection. Commercial NS1 antigen assays are available in Indonesia with variable sensitivity. In an attempt to develop an NS1-based diagnostic test, we successfully cloned NS1 gene of DENV2 to a glutathione Stransferase- based vector pGEX6P-1 in Escherichia coli system. The recombinant protein (pG2NS12) was expressed in E. coli BL21. After induction with isopropyl-β-D-thiogalactoside 0.1 mM for 4 h at 25 °C a recombinant protein GST-NS1 with molecular size of approximately 75 kDa was  obtained. The fusion protein was insoluble and found in the pellet fraction of the cell lysate. Addition of lysozyme (10 mg mL-1) and DNase-I (7.2 mg mL-1) in the lysis buffer was necessary to collect proteins from the pellet fraction. The proteins in the cell pellet were fractionated through Sephadex-G100 column, and GST-NS1 was further purified with Glutathione-Sepharose 4B beads. To obtain pure recombinant NS1 protein to be used in the immunization of mice, the fusion protein was cut with PreScission Protease® by addition of 0.075% Triton-X 100 was necessary to cut the fusion protein. We found that antibodies that recognized the recombinant NS1 protein and DENV2 virus were produced in mice immunized with purified NS1 protein. Therefore, our recombinant NS1 could be used to produce antibody that is potentially useful for developing diagnostic assay to determine the presence of dengue virus NS1 antigen in patient sera.
Effect of Duck Egg Shell Ash with Fly Ash as Cement Substitution in Geopolymer Concrete Larasati, Deka; Nurtanto, Dwi; Utami, Nanin Meyfa
BERKALA SAINSTEK Vol 10 No 2 (2022)
Publisher : Universitas Jember

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.19184/bst.v10i2.29080

Abstract

The increasing production of cement as a basic material for making concrete is due to the increasing demand for concrete throughout the world. Innovative materials are needed as a substitute for cement to reduce the greenhouse effect, fly ash is pozzolanic which in fact has the potential to replace Portland cement as the main ingredient of concrete, and duck eggshell flour has good prospects to be used as an additional precursor material for geopolymer concrete. This study aims to determine further variations of the proportion of duck egg shells, the mechanical properties of the concrete produced using duck egg shell ash with fly ash. This study used an experimental method with the percentage of duck egg shell ash at 0%, 5%, 10%, and 15% of the total use of cement. Using Na₂SiO₃ and NaOH as activator with a concentration of 14M. At the age of 7 and 28 days with room temperature treatment. This study resulted in duck egg shell ash being able to increase the compressive strength of concrete at the age of 28 days with the percentage of substitution up to 5% with a value of 59.26 MPa, in the porosity test the minimum value was at the age of 28 days at a percentage of 5% 0.403%, and the value of the modulus of elasticity. experienced an increase in value at a percentage of 5% with the result 36071.43 MPa. Duck egg shell ash with a proportion of 5% is the optimum substitution where the compressive strength, porosity and modulus of elasticity tests on geopolymer concrete have linear values.