• KOREAN JOURNAL OF CROP SCIENCE
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• 제42권2호
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• pp.247-253
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• 1997

Development of soybean cultivars with great nodulation and high nitrogen fixation activity, derived mostly from mutagenesis, may decrease inputs of chemical fertilizer nitrogen into the soil-plant system. Soybean seeds (cv. Jangyupkong, Hwanggeumkong, and Geomjungkong 1) were treated with three different levels of EMS (ethyl methanesulfonate) concentration(30, 50, and 70mM). Increasing the doses of EMS resulted in decreased field emergence rate of seeds, whereas it did not increase M$_2$ mutation frequencies. This indicated that the most efficient concentration of EMS was 30mM for generating mutants. Extensive mutagenesis of Sinpaldalkong 2 with 30mM EMS was undertaken to isolate soybean mutants with greater nodulation. Approximately 8, 200 M$_2$ families were screened for greater nodulation on 5 mM nitrate after inoculation with Bradyrhizobium japonicum strain YCK213-KFCC-10728. Mutant SS-2 nodulated more than the wild type. Comparison of supernodulation between SS-2 and two nts mutants(nts 1007 and nts 1116) revealed that SS-2 showed the supernodulation character at an earlier growth stage than the two nts mutants. Further studies should be needed to characterize the difference in timing of nodulation between SS-2 and nts mutants.

• Journal of Korean Neurosurgical Society
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• 제67권3호
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• pp.333-344
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• 2024

Objective : Markers of neuroinflammation during ischemic stroke are well characterized, but additional markers of neural damage are lacking. The study identified associations of behavioral disorders after stroke with histologic neural damage and molecular biological change. Methods : Eight-week-old, 25 g male mice of the C57BL/6J strain were subjected to middle cerebral artery occlusion (MCAO) to induce ischemic stroke. The control group was a healthy wild type (WT), and the experimental group were designed as a low severity MCAO1 and a high severity MCAO2 based on post-stroke neurological scoring. All groups underwent behavioral tests, realtime polymerase chain reaction, triphenyltetrazolium chloride (TTC) staining and Hematoxylin and Eosin staining. One-way analysis of variance was used to analyze statistical significance between groups. Results : In TTC staining, MCAO1 showed 29.02% and MCAO2 showed 38.94% infarct volume (p<0.0001). The pro-inflammatory cytokine interleukin (IL)-1β was most highly expressed in MCAO2 (WT 0.44 vs. MCAO1 2.69 vs. MCAO2 5.02, p<0.0001). From the distance to target in the Barnes maze test, WT had a distance of 178 cm, MCAO1 had a distance of 276 cm, and MCAO2 had a distance of 1051 (p=0.0015). The latency to target was 13.3 seconds for WT, 27.9 seconds for MCAO1, and 87.9 seconds for MCAO2 (p=0.0007). Prospero homeobox 1 (Prox1) was most highly expressed in MCAO2 (p=0.0004). Doublecortin (Dcx) was most highly expressed in MCAO2 (p<0.0001). Conclusion : The study demonstrated that histological damage to neural cells and changes in brain mRNA expression were associated with behavioral impairment after ischemic stroke. Prox1 and Dcx may be biomarkers of neural damage associated with long-term cognitive decline, and increased expression at the mRNA level was consistent with neural damage and long-term cognitive dysfunction.

• Korean Journal of Agricultural Science
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• 제5권1호
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• pp.35-41
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• 1978

Extensive selection works on wild yeasts of fruits were carried cut to obtain strains which are applicable to apple wine making. Among the total number of 1,358 yeast strains which were isolated from various fruit samples collected from the vicinity of Daejeon and other regions cf Korea, the strains SH-49, SH-129 and SH-338 were found to be useful. Then experiments on their morphological and physiological characteristics, and on the aspects of practical use in apple wine making were proceeded. The results obtained were as follows: 1. The strains SH-49 and SH-129, particularly SH-49, were appeared to have good fermentation ability, tolerance to sulfur dioxide and to produce fine quality of apple wine. 2. Apple wines made by using the strain, SH-49 and SH-129 contained less amount of total acids than those by other strains. 3. Apple wines of SH-49 and SH-129 were clarified rapidly during the primary fermentation period, and their absorbancy at 430 nm after 45 days of storage were approximately half of others. 4. Apple wine of SH-338 contained higher amount of residual sugar and its quality was superior to others. It is considered that this strain could be used in the production of apple wine of a characteristic quality. 5. The strains SH-49 and SH-338 were identified as a Saccharomyces cerevisiae according to Taxanomic Study of Yeasts by Lodder, however, classification of SH-129 was suspended for further study.

• Korean Journal of Microbiology
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• 제41권2호
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• pp.99-104
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• 2005

Corynebacterial clones which exert regulatory effects on the expression of the glyoxylate bypass genes were isolated using a reporter plasmid carrying the enteric lacZ fused to the aceB promoter of Corynebacterium glutamicum. Some clones carried common fragments as turned out by DNA mapping technique. Subcloning analysis followed by the measurement of $\beta-galactosidase$ activity in Escherichia coli identified the region responsible for the aceB-repressing activity. Sequence analysis of the DNA fragment identified two independent ORFs of ORF1 and ORF2. Among them, ORF2 was turned out to be responsible for the aceB-repressing activity. ORF1 encoded a 23,216 Da protein composed of 206 amino acids. Sequence similarity search indicated that the ORF may encode a ECF-type $\sigma$ factor and designated sigH. To identify the function of sigH, C. glutamicum sigH mutant was constructed by gene disruption technique and the sigH mutant showed growth retardation as compared to the wild type strain. In addition, the mutant strain showed sensitivity to oxidative-stress generating agent plumbagin. This result imply that sigH is probably involved in the stress response occurring during normal cell growth.

• Korean Journal of Microbiology
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• 제43권4호
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• pp.243-249
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• 2007

The centromere is a highly differentiated structure of the chromosome that fulfills a multitude of essential mitotic and meiotic functions. Alphoid DNA (${\alpha}$-satellite) is the most abundant family of repeated DNA found at the centromere of all human chromosomes, and chromosomes of primates in general. The most important parts in the development of Human Artificial Chromosomes (HACs), are the isolation and maintenance of stability of centromeric region. For isolation of this region, we could use the targeting hook with alphoid DNA repeat and cloned by Transformation-Associated Recombination (TAR) cloning technique in yeast Saccharomyces cerevisiae. The method includes rolling-circle amplification (RCA) of repeats in vitro to 5 kb-length and elongation of the RCA products by homologous recombination in yeast. Four types of $35\;kb{\sim}50\;kb$ of centromeric DNA repeat arrays (2, 4, 5, 6 mer) are used to examine the stability of repeats in homologous recombination mutant strains (rad51, rad52, and rad54). Following the transformation into wild type, rad51 and rad54 mutant strains, there were frequent changes in inserted size. A rad52 mutant strain showed extremely low transformation frequency, but increased stability of centromeric DNA repeat arrays at least 3 times higher than other strains. Based on these results, the incidence of large mutations could be reduced using a rad52 mutant strain in maintenance of centromeric DNA repeat arrays. This genetic method may use more general application in the maintenance of tandem repeats in construction of HAC.

• Journal of Life Science
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• 제22권6호
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• pp.837-843
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• 2012

This study was carried out to investigate the morphological and physiological characteristics of six new cultivars of Hypsizygus marmoreus (Hm) and measure endo-, exo-cellular enzyme-specific activity. The domestic wild stain (Hm3-10) and commercial strain in Japan (Hm1-1) were mated by crossing monokaryon mycelia. We gained 58 strains from one of 400 crosses through the $1^{st}$ cultivation experiment, and selected six strains from one of 58 strains through the $2^{nd}$ cultivation experiment. When six of the selected new strains were grown during several spawn culture periods (60, 70, 80, 90, and 100 days), a spawn culture period of more 80 days was considered to be excellent as being shorter than 19~20 days. Therefore, we determined the period of spawn culture as 80 days. Three strains such as Hm15-3, Hm15-4, and Hm17-5 showed an excellent result. When endo-cellular enzyme activity measured eight strains, we obtained a result of that specific activity of ${\alpha}$-amylase at the highest as 73.9~102.2 unit/mg protein, and chitinase is lower than ${\alpha}$-amylase at 8.1~13.1 unit/mg protein. When exo-cellular enzyme activity measured eight strains, we determined the result of that specific activity of ${\alpha}$-amylase is the highest at 5,292~1,184 unit/mg protein, and CMCase and xylanase were 1,140~245 unit/mg protein, 94~575 unit/mg protein, compared to each other. However, the enzyme activity of ${\beta}$-glucosidase and chitinase is low.

• Korean Journal of Soil Science and Fertilizer
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• 제26권4호
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• pp.271-277
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• 1993

An antagonistic bacteria was isolated from rhiaosphere of pepper and corn and identified as Bacillus (B.) subtilis. These B. subtilis B-5 was transformed and marked with the plasmid pCPP4 which possess neomycine resistan. gene. The marked stranins showed growth inhibition to Rhizoctonia (R.) solani, Fusarium (F.) solani, and F. oxysporum in vitro, and were used in studying growth promoting effects on sesame and cabbage. All the identified strains utilize glucose, sucrose, fructose, lactose, mannitol and sorbitol as carbon source, but not rhamnose, and the marked strains also showed characteristics similar to wild-type strains. Germination rate of chinese cabbage and sesame seeds was increased by about 10% or more in the plot to which these strains were inoculated and the effect was higher in soil than in petri dish. The early growth promoting effects of these strains appeared higher, as compared with control plot, in the plots to which B. subtilis B-5 and pathogenic fungi was inoculated together. When the marked strains, B. subtilis B-5NEOr, were inoculated in the rhizosphere of chinese cabbage and sesame with $1.1{\times}10^8CFU/g$ dry soil, the number of inoculated strain was decreased slowly to the level of $10^5{\sim}10^6CFU/g$ dry soil after 4 weeks and the number of Pseudomonas spp. maintanied the level of $10^5CFU/g$ dry soil throught total period, but the number of fungi was decreased rapidly from the early level of $10^8CFU/g$ dry soil to $10^3CFU/g$ dry soil after 4 weeks.

• Research in Plant Disease
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• 제19권4호
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• pp.254-258
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• 2013

Fusaric acid (FA) is a mycotoxin produced by Fusarium species. Its toxicity is relatively low but often associated with other mycotoxins, thus enhancing total toxicity. To date, biosynthetic genes or enzymes for FA have not been identified in F. oxysporum. In order to explore the genetic element(s) for FA biosynthesis, restriction enzyme mediated integration (REMI) procedure as an insertional mutagenesis was employed using FA producing-F. oxysporum strains. Genetic transformation of two F. oxysporum strains by REMI yielded more than 7,100 transformants with efficiency of average 3.2 transformants/${\mu}g$ DNA. To develop a screening system using phytotoxicity of FA, eleven various grains and vegetable seeds were tested for germination in cultures containing FA: Kimchi cabbage seed was selected as the most sensitive host. Screening for FA non-producer of F. oxysporum was done by growing each fungal REMI transformant in Czapek-Dox broth for 3 weeks at $25^{\circ}C$ then observing if the Kimchi cabbage seeds germinated in the culture filtrate. Of more than 5,000 REMI transformants screened, fifty-three made the seeds germinated, indicating that they produced little or fewer FA. Among them, twenty-six were analyzed for FA production by HPLC and two turned out to produce less than 1% of FA produced by a wild type strain. Sequencing of genomic DNA regions (252 bp) flanking the vector insertion site revealed an uncharacterized genomic region homologous (93%) to the F. fujikuroi genome. Further study is necessary to determine if the vector insertion sites in FA-deficient mutants are associated with FA production.

• Journal of the Korean Society of Food Science and Nutrition
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• 제42권10호
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• pp.1690-1700
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• 2013

The objective of this study is to develop and validate predictive growth models for Bacillus cereus (diarrhea type) vegetative cells, spores and Staphylococcus aureus in preprocessed Namul (bracken and Chwinamul) and root vegetables (bellflower and burdock). For validation of model performance, growth data for S. aureus in preprocessed vegetables were collected at independent temperatures (18 and $30^{\circ}C$) not used in the model development. In addition, model performance of B. cereus (diarrhea type) in preprocessed vegetables was validated with an emetic type of B. cereus strain. In primary models, the specific growth rate (SGR) of the B. cereus spores was faster than that of the B. cereus vegetative cells, regardless of the kinds of vegetables at 24 and $35^{\circ}C$, while lag time (LT) of the B. cereus spores was longer than that of the B. cereus vegetative cells, except for burdock. The growth of B. cereus and S. aureus was not observed in bracken at temperatures lower than 13 and $8^{\circ}C$, respectively. The LT models for B. cereus (diarrhea type) in this study were suitable in predicting the growth of B. cereus (emetic type) on burdock and Chwinamul. On the other hand, SGR models for B. cereus (diarrhea type) were suitable for predicting the growth of B. cereus (emetic type) on all preprocessed vegetables. The developed models can be used to predict the risk of B. cereus and S. aureus in preprocessed Namul and root vegetables at the retail markets.

• The Korean Journal of Food And Nutrition
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• 제15권2호
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• pp.112-118
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• 2002

Sporulation in the fission yeast Schizosaccharomyces pombe has been regarded as an important model of cellular development and differentiation. S. pombe cells proliferate by mitosis and binary fission on growth medium. Deprivation of nutrients especially nitrogen sources, causes the cessation of mitosis and initiates sexual reproduction by matting between two sexually compatible cell types. Meiosis is then followed in a diploid cell in the absence of nitrogen source. DNA fragment complemented with the mutations of sporulation gene was isolated from the S. pombe gene library constructed in the vector, pDB 248' and designated as pDB(spo5)1. We futher analyzed six recombinant plasmids, pDB(spo5)2, pDB(spo5)3, pDB(spo5)4, pDB(spo5)5, pDB (spo5)6, pDB(spo5)7 and found each of these plasmids is able to rescue the spo5-2, spo5-3, spo5-4, spo5-5, spo5-6, spo5-7 mutations, respectively. Mapping of the integrated plasmid into the homologous site of the S. pombe chromosomes demonstrated that pDB(spo5)1, and pDB(spu5)Rl contained the spo5 gene. Transcripts of spo5 gene were analyzed by Northern hybridization. Two transcripts of 3.2 kb and 2.5kb were detected with 5kb Hind Ⅲ fragment containing a part of the spo5 gene as a probe. The small mRNA(2.5kb) appeared only when a wild-type strain was cultured in the absence of nitrogen source in which condition the large mRNA (3.2kb) was produced constitutively. Appearance of a 2.5kb spo5-mRNA depends upon the function of the meil, mei2 and mei3 genes.