• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.101-111
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• 2017

The task of improving a fungal strain is highly time-consuming due to the requirement of a large number of flasks in order to obtain a library with enough diversity. In addition, fermentations (particularly those for fungal cells) are typically performed in high-volume (100-250 ml) shake-flasks. In this study, for large and rapid screening of itaconic acid (IA) high-yielding mutants of Aspergillus terreus, a miniaturized culture method was developed using 12-well and 24-well microtiter plates (MTPs, working volume = 1-2 ml). These miniaturized MTP fermentations were successful, only when highly filamentous forms were induced in the growth cultures. Under these conditions, loose-pelleted morphologies of optimum sizes (less than 0.5 mm in diameter) were casually induced in the MTP production cultures, which turned out to be the prerequisite for the active IA biosynthesis by the mutated strains in the miniaturized fermentations. Another crucial factor for successful MTP fermentation was to supply an optimal amount of dissolved oxygen into the fermentation broth through increasing the agitation speed (240 rpm) and reducing the working volume (1 ml) of each 24-well microtiter plate. Notably, almost identical fermentation physiologies resulted in the 250 ml shake-flasks, as well as in the 12-well and 24-well MTP cultures conducted under the respective optimum conditions, as expressed in terms of the distribution of IA productivity of each mutant. These results reveal that MTP cultures could be considered as viable alternatives for the labor-intensive shake-flask fermentations even for filamentous fungal cells, leading to the rapid development of IA high-yield mutant strains.

• Proceedings of the Botanical Society of Korea Conference
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• 1994.09a
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• pp.41-51
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• 1994

Tomato (Lycopersicon esculentum Mill.) has been chosen as a model species for the study of hotomorphogenesis. The aurea (au) and yellow-green-2 (yg-2) mutants which are severely photochrome deficient appear to be phytochrome chromophore mutants. Mutants modified with respect to specific members of the phytochrome gene family: the far-red light-insensitive mutant (fri, for phytochrome A) and the temporarily red light-insensitive mutant (tri, for phytochrome B1) have been identified. Mutants that exhibit an exaggerated phytochrome response are putative transduction-chain mutants affecting an amplification step in phytochrome signal transduction. These mutants are being used to understand the complexities of juvenile anthocyanin in the hypocotyl during seedling de-etiolation.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.50 no.2
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• pp.125-130
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• 2005

In the absence of exogeneous nitrogen supply, evaluation of a symbiosis effectiveness of Bradyrhizobium japonicum USDA 110 in a supernodulating soybean mutant, SS2-2, its wild type, Sinpaldalkong 2, and control genotype, Jangyeobkong, was conducted in this study. Nodules in SS2-2 were initially white and similar to its wild type, Sinpaldalkong 2. At the late stage, the wild type nodules became dark pinkish by maturation, by contrast, mature nodules in SS2-2 remained light green to pinkish, indicating a lack of leghemoglobin. Tap root length was short in nodulated symbiotic SS2-2 than that of its wild type and the control genotype. Nodulated root length and nodule density on root length were significantly increased by B. japonicum inoculation, but no significant increase was observed on root length and percentage of nodulation to total root length. Regardless of Bradyrhizobium inoculation, SS2-2 showed higher nodule dry weight and higher acetylene reduction activity (ARA) when compared with its wild type and the control genotype. Inoculation of B. japonicum leaded the increase of ARA in 47 days after planting (DAP), in part because of nodule development. Supernodulating mutant, SS2-2, less responded to B. japonicum induction in terms of nitrogen fixation and nodulation characteristics than its wild type. Thus, interaction of supernodulating soybean mutant with Bradyrhizobium had less symbiotically associated response than normal nodulating soybean.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2021.04a
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• pp.27-27
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• 2021

Yacon [Samallanthus sonchifolius (Poepp. & Endl.) H. Robinson], a member of Compositae plants, has sweet taste and crisp texture. Unlike other Andean root crops such as potato and sweet potato, the cultivation area of yacon has increased recently, since it is known to have large content of fructooligosaccharides (FOS). Since there are no yacon varieties bred in Korea, we have been trying to create new genetic resources using gamma-ray. The optimal gamma-ray dosage for mutation breeding in yacon was investigated. Crown bud and green bud of yacon were exposed to doses of gamma rays from 20 Gy to 80 Gy, and subsequently planted in a greenhouse. After 50 days of sowing, the survival rates and growth decreased rapidly at doses above 40 Gy, while all of crown bud individuals died above 60 Gy. The median lethal dose (LD50) of crown bud and green bud was 22.4 and 36.6 Gy, and the median reduction doses (RD50) for plant height, fresh weights, and tuberous root weight were 20-40 Gy, respectively. A dose of 20-40 Gy was found to be optimal for mutation breeding in yacon. Considering the growth factors, the optimum doses were determined to be within the range of 20-40 Gy for the selection of useful mutant lines. M2-M3 mutant lines were obtained from 20-60 Gy gamma-ray-irradiated M1 plants through clonal propagation. These mutant lines will be used for the development of a new variety of yacon plant with high FOS and no crack tuberous root.

• Proceedings of the Korean Society of Crop Science Conference
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• 1997.10a
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• pp.38-39
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• 1997

• Bulletin of the Korean Chemical Society
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• v.26 no.3
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• pp.413-417
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• 2005

Fluorescence of green fluorescent protein mutant, 2-5 GFP is observed during denaturation by guanidine. The fluorescence intensity decreases exponentially but the fluorescence lifetime does not change during denaturation. The fluorescence lifetime of the denatured protein is shorter than that of native form. As the protein structure is modified by guanidine, solvent water molecules penetrate into the protein barrel and protonate the chromophore to quench fluorescence. Most fluorescence quenchers do not affect the fluorescence of native form but accelerate the fluorescence intensity decay during denaturation. Based on the observations, a simple model is suggested for the structural change of the protein molecule during denaturation.

• The Plant Pathology Journal
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• v.22 no.4
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• pp.353-359
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• 2006

Transgenic Nicotiana benthamiana plants harboring the coat protein(CP) gene of Zucchini green mottle mosaic virus(ZGMMV) were chosen as a model host for the environmental risk assessment of genetically modified plants with virus resistance. This study was focused on whether new virus type may arise during serial inoculation of one point CP mutant of ZGMMV on the transgenic plants. In vitro transcripts derived from the non-functional CP mutant were inoculated onto the virus-tolerant and -susceptible transgenic N. benthamiana plants. Any notable viral symptoms that could arise on the inoculated transgenic host plants were not detected, even though the inoculation experiment was repeated a total of ten times. This result suggests that potential risk associated with the CP-expressiing transgenic plants may not be significant. However, cautions must be taken as it does not guarantee environmental safety of these CP-mediated virus-resistant plants, considering the limited number of the transgenic plants tested in this study. Further study at a larger scale is needed to evaluate the environmental risk that might be associated with the CP-mediated virus resistant plant.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.5
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• pp.311-315
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• 2002

By the in vitro combinatorial mutagenesis, which is a sequential reaction of PCR mutagenesis and in vitro coupled transcription/translation with Escherichia coli S30 extract, S65 and E222 of green fluorescent protein of Aequarea victoria were comprehensively changed to all possible combinations of amino acids, thus totally 400 mutant (including a wild type) proteins were simultaneously produced and their fluorescent properties were analyzed. Although a few mutations had been reported so far at the 222nd position, replacement E222 to all other19 amino acids gave fluorescent signal to the mutants by changing Ser 65 to Ala together. Among the mutants, replacement to G, A, S, Q, H and C gave relatively high fluorescence. The in vitro combinatorial mutagenesis, therefore, has been proved valuable for comprehensive structure-function studies of proteins.

• Journal of Microbiology and Biotechnology
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• v.20 no.3
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• pp.460-466
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• 2010

A mutant of green fluorescent protein ($GFPmut3^*$) from the jellyfish Aequorea victoria was cyclized in vitro and in vivo by the use of a naturally split intein from the dnaE gene of Synechocystis species PCC6803 (Ssp). Cyclization of $GFPmut3^*$ was confirmed by amino acid sequencing and resulted in an increased electrophoretic mobility compared with the linear $GFPmut3^*$. The circular $GFPmut3^*$ was $5^{\circ}C$ more thermostable than the linear form and significantly more resistant to proteolysis of exopeptidase. The circular $GFPmut3^*$ also displayed increased relative fluorescence intensity. In addition, chemical stability of $GFPmut3^*$ against GdnHCl revealed more stability of the circular form compared with the linear form.

• Korean Journal of Medicinal Crop Science
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• v.8 no.4
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• pp.334-341
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• 2000

This study was conducted to determine the effects and optimum concentrantion of chemical mutagens, colchicine, EMS (ethyl methan sulfonate), MNU (1-methyl-3-1­-nitrosoguanidinenitro), sodium azide $(NaN_3)$ for induction of mutant plants. In order to induce the mutants of Dianthus superbus L, immature seed were pre-soaked in the warter adding each mutagens and concentration of EMS, colchicine, MNU, and sodium azide $(NaN_3)$. Comparision of morphological characteristic and seed germination in each mutant plants differed depending on mutagen sources and their concentrations. When 0.2% EMS were treated on seed, germination decreased to 12% while untreated control was germinated 76.6% for twenty days. Treatments of colchicine appeared higher germination than other mutagen but not survived. The survival rate was extremely decreased in MNU treatment at 0.5mM and chlorophyll-mutant plantlets were obtained by sodium azide treatment at 0.2mM. Chlorophyll mutants were produced by pre-soaking the immature seed of Dianthus superbus L. with mutagen, sodium azide. The control plants appeared normal green leaf color, while mutant plant after mutagenic treatment of immature seed results in yellow­-green stripes and albino in normal green leaf tissue. RAPD was carried out to check the genetic modification of regenerated plants by mutagen treatments at 0.2mM sodium azide. Three polymorphic DNA fragments out of thirty-seven obtained by RAPDs were observed in regenerated plants using five decamer primers.