• Journal of Life Science
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• v.28 no.12
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• pp.1416-1423
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• 2018

With strong biotin binding affinity ($K_D=10^{-14}M$), the tetrameric feature of streptavidin could be used to increase the antigen binding activity of a camel heavy chain (VHH) antibody through their fusion, here stained with biotinylated horseradish peroxidase and subsequent immunoassays ELISA and Western blot analysis. For this application, we cloned the streptavidin gene amplified from the Streptomyces avidinii chromosome by PCR, and this was fused to the gene of the 8B9 VHH antibody which is specific to green fluorescent protein (GFP) antigens. To express a soluble fusion protein in Escherichia coli, we used the pUC119 plasmid-based expression system which uses the lacZ promoter for induction by IPTG, the pelB leader sequence at the N-terminus for secretion into the periplasmic space, and six polyhistidine tags at the C-terminus for purification of the expressed proteins using an $Ni^+$-NTA-agarose column. Although streptavidin is toxic to E. coli because of its strong biotin binding property, this soluble fusion protein was expressed successfully. In SDS-PAGE, the size of the purified fusion protein was 122.4 kDa in its native condition and 30.6 kDa once denatured by boiling, suggesting the tetramerization of the monomeric subunit by non-covalent association through the streptavidin moiety fusing to the 8B9 VHH antibody. In addition, this fusion protein showed biotin binding activity similar to streptavidin as well as GFP antigen binding activity through both ELISA and Western blot analysis. In conclusion, the protein resulting from the fusion of an 8B9 VHH antibody with streptavidin was successfully expressed and purified as a soluble tetramer in E. coli; it showed both biotin and GFP antigen binding activity suggesting the possible production of a tetrameric and bifunctional VHH antibody.

• Journal of Korean Society of Forest Science
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• v.71 no.1
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• pp.14-21
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• 1985

This study was carried out in order to prevent the devaluation of wood itself and wood products causing by anisotropy, hygroscopicity, shrinkage and swelling - properties that wood itself only have, in order to improve utility of wood, by emphasizing the natural beautiful figures of wood, to develop the dimensional stabilization techniques of wood with PEG that it is a cheap, non-toxic and the impregnation treatment is not difficult, on the effects of PEG molecular weights (200, 400, 600, 1000, 1500, 2000, 4000, 6000) and species (Pinus densiflora S. et Z., Larix leptolepis Gordon., Cryptomeria japonica D. Don., Cornus controversa Hemsl., Quercus variabilis Blume., Prunus sargentii Rehder.). The results were as follows; 1) PEG loading showed the maximum value (137.22%, Pinus densiflora, in PEG 400), the others showed that relatively slow decrease. The lower specific gravity, the more polymer loading. 2) Bulking coefficient didn't particularly show the correlation with specific gravity, for the most part, indicated the maximum values in PEG 600, except that the bulking coefficient of Quercus variabilis distributed between the range of 12-18% in PEG 400-2000. In general, the bulking coefficient of hardwood was higher than that of softwood. 3) Although there was more or less an exception according to species, volumetric swelling reduction was the greatest in PEG 400. That is, its value of Cryptomeria japonica was the greatest value with 95.0%, the others indicated more than 80% except for Prunus sargentii, while volumetric swelling reduction was decreased less than 70% as the molecular weight increase more than 1000. 4) The relative effectiveness of hardwood with high specific gravity was outstandingly higher than softwood. In general, the relative effectiveness of low molecular weight PEG was superior to those of high molecular weight PEG except that Quercus variabilis showed more than 1.6 to the total molecular weight range, while it was no significant difference as the molecular weight increase more than 4000. 5) According to the analysis of the results mentioned above, the dimensional stabilization of hardwood was more effective than softwood. Although volumetric swelling reduction was the greatest at a molecular weight of 400. In the view of polymer loading, bulking coefficiency reduction of swelling and relative effectiveness, it is desirable to use the mixture of PEG of molecular weight in the range of 200-1500. To practical use, it is recommended to study about the effects on the mixed ratio on the bulking coefficient, reduction of swelling and relative effectiveness.

• Korean Journal of Environmental Agriculture
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• v.23 no.4
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• pp.203-210
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• 2004

This study was conducted to assessment organic compound and bioassay (density of inhabited animal, fluctuation of predominant fungi, and survival ratio of earthworm) for finding damage on red pepper by heavily amount application of sludges in soil, which was treated with 3 pharmaceutical byproducts and a cosmetic industry wastewater sludge as raw materials of compost, and for establishing estimation method. HEM contents in the soil treated with pharmaceutical byproducts sludge2 (PS2) and cosmetic sludge (CS) were 0.51, 1.10 mg/kg respectively. PAHs content of PS2 treatment in the soil was 3406.8 ug/kg on July 8. In abundance of soil faunas, the pharmaceutical byproducts sludge2 treatment was the most highest. The next was decreased in the order of pig manure (PM) and the cosmetic sludge treatment. However the other pharmaceutical sludge treatments were remarkably reduced populations of soil inhabited animals. In upland soil treated with organic sludges, the numbers of bacteria and fungi of the pharmaceutical sludge treatment were 736, 909 cfu/g and those of the cosmetic sludge treatment were 440, 236 cfu/g, respectively. The pharmaceutical sludge treatments and the cosmetic sludge treatment in identification of predominant bacteria were not any tendency to compare with non fertilizer and pig manure treatments, but they had diverse bacteria than NPK treatment. In microcosm tests, the survival of the tiger earthworm in five soil samples was hardly affected against the soil of PSI (20%) after three months treated in the upland But after six months, survival of PS1 was 80%. At present, raw material of compost was authorized by contents of organic matter, heavy metal (8 elements), and product processing according to 'The specified gist on possible materials of using after analysis and investigation among raw materials of compost', however, for preparing to change regulation of raw material of compost and for considering to possibility of application, this study was conducted to investigate toxic organic compound and bioassay methods using inhabited animal, fungi, and earthworm without current regulation.

• Korean Journal of Soil Science and Fertilizer
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• v.2 no.1
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• pp.53-68
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• 1969

The nutriophysiological response of rice plant to root environment was investigated with eye observation of root development and rhizosphere in situation. The results may be summarized as follows: 1) The quick decomposition of organic matter, added in low yield soil, caused that the origainal organic matter content was reached very quickly, in spite of it low value. In high yield soil the reverse was seen. 2) In low yield soil root development, root activity and T/R value were very low, whereas addition of organic matter lowered them still wore. This might be contributed to gas bubbles around the root by the decomposition of organic matter. 3) Varietal difference in the response to root environment was clear. Suwon 82 was more susceptible to growth-inhibitine conditions on low-yield soil than Norin 25. 4) Potassium uptake was mostly hindered by organic matter, while some factors in soil hindered mostly posphorus uptake. When the organic matter was added to such soil, the effect of them resulted in multiple interaction. 5) The root activity showed a correlation coeffieient of 0.839, 0.834 and 0.948 at 1% level with the number of root, yield of aerial part and root yield, respectively. At 5% level the root-activity showed correlation-coefficient of 0.751, 0.670 and 0.769 with the uptake of the aerial part of respectively. N, P and K and a correlation-coefficient of 0.729, 0.742 and 0.815 with the uptake of the root of respectively N.P. and K. So especially for K-uptake a high correlation with the root-activity was found. 6) The nitrogen content of the roots in low-yield soil was higher than in high-yield soil, while the content in the upper part showed the reverse. It may suggest ammonium toxicity in the root. In low-yield soil Potassium and Phosphorus content was low in both the root and aerial part, and in the latter particularly in the culm and leaf sheath. 7) The content of reducing sugar, non-recuding sugar, starh and eugar, total carbohydrates in the aerial part of plants in low yield soil was higher than in high yield soil. The content of them, especially of reducing sugar in the roots was lower. It may be caused by abnormal metabolic consumption of sugar in the root. 8) Sulfur content was very high in the aerial part, especially in leaf blade of plants on low yield soil and $P_2O_5/S$ value of the leaf blade was one fifth of that in high yield soil. It suggests a possible toxic effect of sulfate ion on photophosphorization. 9) The high value of $Fe/P_2O_5$ of the aerial part of plants in low yield soil suggests the possible formation of solid $Fe/PO_4$ as a mechanical hindrance for the translocation of nutrients. 10) Translocation of nutrients in the plant was very poor and most nutrients were accumulated in the root in low yield soil. That might contributed to the lack of energy sources and mechanical hindrance. 11) The amount of roots in high yield soil, was greater than that in low yield soil. The in high-yield soil was deep, distribution of the roots whereas in the low-yield soil the root-distribution was mainly in the top-layer. Without application of Nitrogen fertilizer the roots were mainly distributed in the upper 7cm. of topsoil. With 120 kg N/ha. root were more concentrated in the layer between 7cm. and 14cm. depth. The amount of roots increased with the amount of fertilizer applied.