• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.62-65
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• 2002

We have previously shown that the addition of silkworm hemolymph to a culture medium increases the longevity of insect and mammalian cells by inhibiting apoptosis. This indicates that the component which inhibits apoptosis is contained in the silkworm hemolymph, The apoptosis-inhibiting component was isolated from silkwonn hemolymph and characterized in our previous study. A database search using the N-terminal amino acid sequence of this component as a template resulted in a 95% homology with a low molecular weight lipoprotein, the so called ’30K protein' of unknown function. In this study, the 30K protein gene was expressed in mammalian and insect cells to confirm the apoptosis-inhibiting effect. The overexpression of 30K protein in mammalian cell inhibited the staurosporin-induced apoptosis by the prevention of the activation of caspase 3. Using an Autographa californicanuclear polyhedrosis virus (AcNPV) system, the 30K protein was overexpressed also in insect cells. The expression of the 30K protein increased the longevity of baculovirus-infected insect cells by inhibiting apoptosis. These results suggest that the 30K protein is a novel anti-apoptotic protein.

• Korean journal of applied entomology
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• v.34 no.3
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• pp.218-223
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• 1995

A nuclear polyhedrosis virus isolated from the oriental tobacco budworm larvae, Helicoverpa assulta (Guenee) was characterized by electron microscopy, SDS-PAGE, restriction endonuclease analysis and cross infectivity. The shape of a polyhedron was $1.0\mu\textrm{m}$ in average with icosahdral outline, and the virus particle was $65nm\times300nm$ in average with rod-shape. The nuclear polyhedrosis virus was contained a single nucleocapsid within a viral envelope embedded in a polyhedron. The polyhedral protein was composed of a single polypeptide with a M.W. of 31 Kd. The genome size of the virus by restriction endonuclease analysis was about 120 Kb. Among several nuclear polyhedrosis viruses, the nuclear polyhedrosis virus from Helicoverpa assulta (HaNPV) and Autographa california nuclear polyhedrosis virus (AcNPV) were infected the oriental tobacco budworm larvae.

• Biomedical Science Letters
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• v.9 no.4
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• pp.177-181
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• 2003

The baculovirus/Spodoptera frugiperda (Sf) cell system has become popular for the production of large amounts of the human erythrocyte glucose transporter, GLUT1, heterologously. However, it was not possible to show that the expressed transporter in insect cells could actually transport glucose. The possible reason for this was that the activity of the endogenous insect glucose transporter was extremely high and so rendered transport activity resulting from the expression of exogenous transporter very difficult to detect. Sf21-AE cells are commonly employed as the host permissive cell line to support the baculovirus AcNPV replication and protein synthesis. The cells grow well on TC-100 medium that contains 0.1 % D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, unlike the human glucose transporter, very little is known about properties of the endogenous sugar transporter(s) in insect cells. Thus, the uptake of 2-deoxy-D-glucose (2dGlc) by Sf21-AE cells and the inhibition of 2dGlc transport in the insect cells by fructose and cytochalasin B were investigated in the present work. The binding assay of cytochalasin B was also performed, which could be used as a functional assay for the endogenous glucose transporter(s) in the insect cells. Sf21-AE cells were infected with the recombinant virus AcNPV-GT or no virus, at a multiplicity of infection (MOI) of 5. Infected cells were resuspended in PBS plus and minus 300 mM fructose, and plus and minus 20 $\mu$M cytochalasin B for use in transport assays. Uptake was measured at 28$^{\circ}C$ for 1 min, with final concentration of 1 mM deoxy-D-glucose, 2-[1,2-$^3$H]- or glucose, L-[l,$^3$H]-, used at a specific radioactivity of 4 Ci/mol. The results obtained demonstrated that the sugar uptake in uninfected cells was stereospecific, and was strongly inhibited by fructose but only poorly inhibitable by cytochalasin B. It is therefore suggested that the Sf21-AE glucose transporter has very low affinity for cytochalasin B, a potent inhibitor of human erythrocyte glucose transporter.

• Journal of Microbiology and Biotechnology
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• v.11 no.5
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• pp.853-857
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• 2001

The effect of silkworm hemolymph on the inhibition of baculovirus-induced insect cell apoptosis was quantitatively investigated using a flow cytometric analysis. Spodoptera frugiperda (Sf9) cell and Autographa californica nuclear polyhedrous virus (AcNPV) were used as a model for insect cell and baculovirus in this study, respectively. Compared with a mammalian cell cycle, the fraction of G1 cells was relatively small in the Sf9 cell cycle. Silkworm hemolymph did not affect the Sf9 cell-cycle distribution before the baculovirus infection. However, the fraction of cells which are not in the sub-G1 phase remained at a high level for 3 days after the infection in the medium without silkworm hemolymph, while it remained at a high level for 7 days after the infection in the medium supplemented with silkworm hemolymph. The fractions of apoptotic cells in the sub-G1 phase were $4.7\%$, and 4 days after infection, $22.7\%$, in the media with and without silkworm hemolymph, respectively.

• Biomedical Science Letters
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• v.11 no.3
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• pp.327-332
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• 2005

Sf21 cells become popular as the host permissive cell line to support the baculovirus AcNPV replication and protein synthesis. The cells grow well on TC-100 medium that contains $0.1\%$ D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, unlike human glucose transporters, very little is known about the characteristics of the endogenoussugar transporter(s) in Sf21 cells. Thus, some kinetic properties of the sugar transport system were investigated, involving the uptake of 2-deoxy-D-glucose (2dG1c). In order to obtain a true measure of the initial rate of uptake, the uptake of $[^3H]2dGlc$ from both low $(100{\mu}M)$ and high (10 mM) extracellular concentrations was measured over periods ranging from 30 sec to30 min. The data obtained indicated that the uptake was linear for at least 2 min at both concentrations, suggesting that measurements made over a 1min time course would reflect initial rates of the jexpse uptake. To determine $K_m\;and\;V_{max}$ of the endogenous glucose transporter(s) in Sf21 cells, the uptake of 2dG1c was measured over a range of substrate concentrations $(50{\mu}M\~10mM)$ 2dG1c uptake by the Sf21 cells appeared to involve both saturable and non-saturable (or very low affinity) components. A saturable transport system for 2dG1c was relatively high, the $K_m$ value for uptake being < 0.45 mM. The $V_{max}$ value obtained for 2dG1c transport in the Sf21 cells was about 9.7-folds higher than that reported for Chinese hamster ovary cells, which contain a GLUT1 homologue. Thus, it appeared that the transport activity of the Sf21 cells was very high. In addition, the Sf21 glucose transporter was found to have very low affinity for cytochalasin B, a potent inhibitor of human erythrocyte glucose transporter

• International Journal of Advanced Culture Technology
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• v.3 no.1
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• pp.120-128
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• 2015

In this paper, Residential roof top photovoltaic system with 9.9 kW design is proposed. The system composed of 200 Watts solar array 33 panels connecting in series 10 strings and parallels 3 strings which have maximum voltage and current are 350 V and 23.8 A. The 10 kW sinusoidal grid-connected inverter with window voltage about 270-350 is selected to convert and transfer DC Power to AC Power at PCC (Point of Common Coupling) of power system following to utility standard. However the impact of fluctuation and uncertainty of weather condition of PV may decrease the voltage stability and voltage collapse of power system. In order to solve this problem the energy storage such 120 V 1200 Ah battery bank and 30 kVAR capacitor are designed for voltage stability control. The other expensed for installing the system such battery charger, cable, accessories and maintenance cost are concerned. The economic analysis by using investment from money loan with interest about 7% and use own money which loss income of deposit about 3% are calculated as 671,844 and 547,044 for PV system with energy storage and non energy storage respectively. The solar energy from PV is about 101,616 Bath per year which evaluated by using the value of $5kWh/m^2/day$ from average peak sun hour (PSH) of the Thailand and 6.96 Bath/kWh of Feed in Tariff Incentive. The payback periods of four scenarios are proposed follow as i) PV system with energy storage and use loan money is 15 years ii) PV system with no energy storage and use loan money is 10 years iii) PV system with energy storage and use deposit money is 9 years iv) PV system with energy storage and use deposit money is 7 years. In addition, the other scenarios of economic analysis such no FIT support and other type of economic analysis such NPV and IRR are proposed in this paper.

• World Technopolis Review
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• v.3 no.2
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• pp.89-96
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• 2014

Center for Research of Science and Technology ("PUSPIPTEK") has 460 hectares land area, still maintained as a green area with more than 30% green space. There are 47 centers for research and testing technology, technology-based industries, and as well as public supporting facilities in PUSPIPTEK area. Based on the concepts developed to make this area as an ecological region, PUSPIPTEK can be seen as a model of eco-innovation. The purpose of this research is to develop a model of water, energy and waste management with eco-innovation concept. As a new approach in addressing environmental degradation and maintaining the sustainability of ecosystem, studies related to eco-innovation policy that combines the management of water, energy and waste in the region has not been done. In order to achieve the objectives of the research, a series of techniques for collecting data on PUSPIPTEK existing conditions will be carried out, which includes utilities data (water, electricity, sewage) and master plan of this area. The savings over the implementation of the concept of eco-innovation in water, energy, and waste management were calculated and analyzed using quatitative methods. The amount of cost savings and feasibility were then calculated. Eco innovation in water management among other innovations include the provision of alternative sources of water, overflow of rain water and water environments utilization, and use of gravity to replace the pumping function. Eco-innovation in energy management innovations include the use of LED and solar cell for air conditioning. Eco-innovation in waste management includes methods of composting for organic waste management. The research results: (1) The savings that can be achieved with the implementation of eco innovation in the water management is Rp. 3,032,640 daily, or Rp.1,106,913,600 annually; (2) The savings derived from the implementation of eco innovation through replacement of central AC to AC LiBr Solar Powered will be saved Rp.1,933,992,990 annually and the use of LED lights in the Public street lighting PUSPIPTEK saved Rp.163,454,433 annually; (3) Application of eco innovation in waste management will be able to raise awareness of the environment by sorting organic, inorganic and plastic waste. Composting and plastic waste obtained from the sale revenue of Rp. 44,016,000 per year; (4) Overall, implementation of the eco-innovation system in PUSPIPTEK area can saves Rp. 3,248,377,023 per year, compared to the existing system; and (5)The savings are obtained with implementation of eco-innovation is considered as income. Analysis of the feasibility of the implementation of eco-innovation in water, energy, and waste management in PUSPIPTEK give NPV at a 15% discount factor in Rp. 3,895,228,761; 23.20% of IRR and 4.48 years of PBP. Thus the model of eco-innovation in the area PUSPIPTEK is feasible to implement.

• Journal of Microbiology and Biotechnology
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• v.16 no.10
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• pp.1583-1590
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• 2006

T4 endonuclease V (T4 endo V) [EC 3. 1. 25. 1], found in bacteriophage T4, is responsible for excision repair of damaged DNA. The enzyme possesses two activities: a cyclobutane pyrimidine dimer DNA glycosylase (CPD glycosylase) and an apyrimidic/apurinic endonuclease (AP lyase). T4 denV (414 bp cDNA) encoding T4 en do V (138 amino acid) was synthesized and expressed using either an expression vector, pTriEx-4, in E. coli or a baculovirus AcNPV vector, pBacPAK8, in insect cells. The recombinant His-Tag/T4 endo V (rHis-Tag/T4 endo V) protein expressed from bacteria was purified using one-step affinity chromatography with a HiTrap Chelating HP column and used to make rabbit anti-His-Tag/T4 endo V polyclonal antibody for detection of recombinant T4 endo V (rT4 endo V) expressed in insect cells. In the meantime, the recombinant baculovirus was obtained by cotransfection of BacPAK6 viral DNA and pBP/T4 endo V in Spodoptera frugiperda (Sf21) insect cells, and used to infect Sf21 cells to overexpress T4 endo V protein. The level of rT4 endo V protein expressed in Sf21 cells was optimized by varying the virus titers and time course of infection. The optimal expression condition was set as follows; infection of the cells at a MOI of 10 and harvest at 96 h post-infection. Under these conditions, we estimated the amount of rT4 endo V produced in the baculovirus expression vector system to be 125 mg/l. The rT4 endo V was purified to homogeneity by a rapid procedure, consisting of ion-exchange, affinity, and reversed phase chromatographies, based on FPLC. The rT4 endo V positively reacted to an antiserum made against rHis-Tag/T4 endo V and showed a residual nicking activity against CPD-containing DNA caused by UV. This is the first report to have T4 endo V expressed in an insect system to exclude the toxic effect of a bacterial expression system, retaining enzymatic activity.