• Korean Journal of Soil Science and Fertilizer
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• v.25 no.4
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• pp.387-393
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• 1992

The fate of inoculum strain of Bradyrhizobium japonicum was studied by using genetically marked strain. RJB6 $str^rnal^rneo^r$. A spontaneous mutant of B. japonicum isolated from nodules was made to have antibiotic resistance against streptomycin and nalidixic acid. In order to make genetically marked strain, neomycine resistant gene(Tn5) was introduced into this spontaneous mutant by conjugation with E. coli containing pSUP2021. The southern hybridization was carried out to confirm the plasmid insertion. Hybridization of chromosome DNA using pSUP2021(Tn5) as a probe showed that Tn5 was located on the 4.9kb fragment of chromosome. Soybean seeds were planted into a soil previously cultivated with soybean and inoculated with different cell densities of marked strain. Fourty days after planting, the inoculation effects on nodule number, nodule fresh weight, plant height and nitrogen content in the plot inoculated with heavy cell suspension was a little better than those in the plot with low inoculation. The recovery percentage of the marked strains was about 12% in the plot inoculated with heavy density cell suspension, while 5% in the plot inoculated with low cell suspension.

• International Journal of Industrial Entomology and Biomaterials
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• v.9 no.2
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• pp.173-181
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• 2004

To delay the onset of apoptosis in the culture, transformed Tn 5B1-4 cells harboring anti-apoptotic genes, bcl-2 and baculovirus p35, have been established and analyzed for their anti-apoptotic ability in suspension culture using spinner flasks. In the suspension culture at agitation speeds of 100 rpm and 200 rpm, the cell growth of cell clone expressing Bcl-2 protein was much higher than other two clones and the maximum cell density of the clone was 6.0 ${\times}$ 10$^{6}$ cells/ml and 6.2 ${\times}$ 10$^{6}$ cells/ml at day three of the incubation. On the other hand, the cell growth of cell clone expressing baculovirus protein P35 was much higher than other two clones in suspension culture at agitation speed of 300 rpm and the maximum cell density of the clone was 6.1 ${\times}$ 10$^{6}$ cells/ml at day three of the incubation. Based on the pattern of genomic DNA laddering and the microscopic observation of apoptotic bodies, the more apoptotic bodies are induced in Tn 5B1-4 control cell clone at higher agitation speed. This result shows that the shear stress can be a main factor in inducing apoptosis in spinner flask culture. At low agitation speed, cell clone expressing Bcl-2 was more effective in delaying the onset of apoptosis than the cell clone expressing P35. On the other hand, at high agitation speed, cell clones expressing baculovirus P35 was more effective in delaying the onset of apoptosis than the cell clone expressing Bcl-2. Therefore, anti-apoptotic genes, bcl-2 and baculovirus p35, can playa distinct role depending on agitation speed in the suspension culture.

• Journal of Plant Biotechnology
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• v.30 no.1
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• pp.97-102
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• 2003

Lettuce (Lactuca sativa) was transformed with Agrobacterium tumefacience LBA4404 containing human granulocyte macrophage colony stimulating factor (hGM-CSF) gene to produce in cell suspension cultures. Cell suspension culture was established using callus from transgenic lettuce plant. Integration of hGM-CSF gene into plant chromosome was confirmed through genomic PCR and Southern blot analysis. In addition, Northern blot analysis indicated the expression of the introduced hGM-CSF gene in transformed lettuce. The recombinant hGM-CSF was expressed in transgenic cell cultures derived from transgenic plants as a yield of about 149.0 $\mu\textrm{g}$/L in culture filtrate, which was determined by ELISA. These results demonstrated that transformed lettuce cell suspension cultures could be used as a production system of therapeutic proteins such as hGM-CSF.

• Transactions of the Korean Society of Automotive Engineers
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• v.12 no.3
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• pp.159-169
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• 2004

In this study, integration control of air-cell seat and semi-active suspension is proposed to minimize the road-tyre force which can cause uncomfortable feeling to rider. The proposed integration control with sliding perturbation observer is consisted of air-cell seat control which uses the force generated by air-cell and the sky-hook control. The air-cell seat itself has been modeled as a 1 degree of freedom spring-damper system. The actual characteristics of the air-cell have been analyzed through experiments. In this paper, we introduces a new robust motion control algorithm using partial state feedback for a nonlinear system with modelling uncertainties and external disturbances. The major contribution of this work is the development and design of robust observer for the state and the perturbation. The combination skyhook controller and air-cell controller using the observer improves control performance, because of the robust routine called Sliding Observer Design for Integration Control of Air-Cell Seat and Semi-active Suspension. The simulation results show a high accuracy and a good performance.

• Proceedings of the Botanical Society of Korea Conference
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• 2002.04a
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• pp.89-89
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• 2002

• Journal of Environmental Health Sciences
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• v.22 no.3
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• pp.98-102
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• 1996

This study investigated performance of the phenol degradation and reaction characteristics according to variation of phenol volumetric loading rates and dilution rates in suspension and PACT reactors using Pseudomonas sp. B3. 1. Removal efficiencies of the PAC unit indicated about 100 % with phenol volumetric loading rates from 0.4 phenol $kg/ma^3\cdot d$ to 1.2 phenol $kg/m^3\cdot d$, however, which of the suspension reactor showed about 100% with from 0.2 phenol $kg/m^3\cdot d$ to 0.75 phenol $kg/ma^3\cdot day$. 2. The cell density slightly was decreased from 298.2 mg/l to 272 mg/l, when dilution rate for suspension was reactor increased from 0.4 to 1.41 1/d, and also the cell density suddenly was decreased to 145.5 mg/l and was washed out at the dilution rate higher than 1.60 1/d. But the cell density for the PAC unit was linearly decreased with dilution rate of from 0.8 to 3.0 1/d, and showed 220.75 mg/l at maximum dilution rate. 3. The phenol utilization rate was increased from 0.008 to 0.031 phenol g/l$\cdot$h, when dilution rate for suspension reactor was increased from 0.4 to 1.5 1/d, however, the rate for the PAC unit was linearly increased from 0.017 to 0.061 phenol g/l$\cdot$h as variation changes from 0.017 to 0.061 phenol g/l$\cdot$h dilution rate.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.6
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• pp.331-334
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• 2002

Culture conductivity and on-line NADH fluorescence were used to measure cellular growth in plant cell suspension cultures of Podophyllum hexandrum. An inverse correlation between dry cell weight and medium conductivity was observed during shake flask cultivation. A linear relationship between dry cell weight and culture NADH fluorescence was obtained during the exponential phase of batch cultivation In a bioreactor under the pH stat (pH 6) conditions. It was observed that conductivity measurement were suitable for biomass characterisation under highly dynamic uncontrolled shake flask cultivation conditions. However, if the acid/alkali feeding is done for pH control the conductivity measurement could not be applied. On the other hand the NADH fluorescence measurement allowed online-in situ biomass monitoring of rather heterogenous plant cell suspension cultures in bioreactor even under the most desirable pH stat conditions.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.3
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• pp.198-204
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• 2005

Seeds of neem were collected from different parts of India and analyzed for their azadirachtin content by High Performance Liquid Chromatography (HPLC). In order to assess the effects of genotypic and geographical variation on azadirachtin content in cell cultures, callus development was attempted in the seeds containing high and low concentration of azadirachtin. The concentration of azadirachtin in callus cultures was significantly affected by the explant source. Seed kernels with higher azadirachtin content produced higher azadirachtin content in callus cultures and lower azadirachtin content was seen in callus cultures produced from seed kernels with low azadirachtin content. The protocol for development of elite stock culture of Azadirachta indica was established with the objective of selecting a high azadirachtin-producing cell line. The highest azadirachtin-producing cell line was selected and the effects of different media and illumination conditions on growth and azadirachtin production were studied in shake flask suspension culture. Detailed batch growth kinetics was also established. These studies provided elite starter culture and associated protocols for cultivation of A. indica plant cell culture in the bioreactor.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.6
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• pp.423-427
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• 2004

A simple purification procedure of bioactive human granulocyte macrophage colony stimulating factor (hGM-CSF) secreted in rice cell suspension culture has previously been described. In this study the protein was purified to apparent homogeneity with an overall yield of $80.1\%$ by ammonium sulfate precipitation and a single chromatographic step involving FPLCanion exchange chromatography. The purified hGM-CSF revealed at least five glycosylated forms ranging from $21.5{\~}29$ kDa, and its biological activity was independent of the glycosylation pattern. This is the first purification report of recombinant hGM-CSF to apparent homogeneity from rice cell suspension cultures.

• Applied Biological Chemistry
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• v.36 no.1
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• pp.1-6
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• 1993

Chitinase was induced in rice cell suspension culture with treatment of chitooligosaccharides mixture. Among eleven isozymes found in 10% polysacrylamide gel electropherogram, four isozymes were identified as induced enzymes. Acidic chitinase fraction separated in DEAE-cellulose column chromatography, includes three induced chitinase, while basic fraction contains only one induced isozyme. Treatment of chitooligosaccharides mixture enhanced the contents in both protein and chitinase activity in cell suspension culture media, but increase in chitinase activity was much higher than in protein.