• Mycobiology
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• v.31 no.1
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• pp.32-35
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• 2003

Restriction enzyme-mediated integration(REMI) was used to transform uracil auxotrophs of Pleurotus ostreatus to prototrophy. When protoplasts of Pleurotus ostreatus were treated by the reaction mixture containing 10 units of BamHI, the frequency of REMI was about 64 transformants per 1 ${\mu}g$ of DNA. This efficiency was increased by 14.2 times compared with that of the conventional PEG transformation. The optimal condition for REMI of P. ostreatus was achieved when 1 ${\mu}g$ of linearized pTRura3-2 DNA was added into $1{\times}10^7$ protoplasts along with 10 units BamHI. Southern blot analysis revealed that about 50% of transformants examined were caused by REMI event and 30% carried single copy insertion at the genome. This suggested that the REMI method might be a useful tool for efficient transformation and tagging mutagenesis of P. ostreatus.

• Geophysics and Geophysical Exploration
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• v.9 no.1
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• pp.108-113
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• 2006

Urban conditions, such as existing underground facilities and ambient noise due to cultural activity, restrict the general application of conventional geophysical techniques. At a tunnelling site in an urban area along an existing railroad, we used the refraction microtremor (REMI) technique (Louie, 2001) as an alternative way to get geotechnical information. The REMI method uses ambient noise recorded by standard refraction equipment and a linear geophone array to derive a shear-wave velocity profile. In the inversion procedure, the Rayleigh wave dispersion curve is picked from a wavefield transformation, and iteratively modelled to get the S-wave velocity structure. The REMI survey was carried out along the line of the planned railway tunnel. At this site vibrations from trains and cars provided strong seismic sources that allowed REMI to be very effective. The objective of the survey was to evaluate the rock mass rating (RMR), using shear-wave velocity information from REMI. First, the relation between uniaxial compressive strength, which is a component of the RMR, and shear-wave velocity from laboratory tests was studied to learn whether shear-wave velocity and RMR are closely related. Then Suspension PS (SPS) logging was performed in selected boreholes along the profile, in order to draw out the quantitative relation between the shear-wave velocity from SPS logging and the RMR determined from inspection of core from the same boreholes. In these tests, shear-wave velocity showed fairly good correlation with RMR. A good relation between shear-wave velocity from REMI and RMR could be obtained, so it is possible to estimate the RMR of the entire profile for use in design of the underground tunnel.

• Research in Plant Disease
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• v.19 no.4
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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.

• 한국지구물리탐사학회:학술대회논문집
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• 2005.05a
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• pp.157-164
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• 2005

Urban conditions such as underground facilities and ambient noises due to cultural activity restrict the application of conventional geophysical techniques in general. We used the refraction microtremor (REMI) technique as an alternative way to get the geotechnical information, in particular shear-wave (S-wave) velocity information, at a site along an existing rail road. The REMI method uses ambient noises recorded using standard refraction equipment to derived shear-wave velocity information at a site. It does a wavefield transformation on the recorded wavefield to produce Rayleigh wave dispersion curve, which are then picked and modeled to get the shear-wave velocity structure. At this site the vibrations from the running trains provided strong noise sources that allowed REMI to be very effective. REMI was performed along the planned new underground rail tunnel. In addition, Suspension PS logging (SPS) were carried out at selected boreholes along the profile in order to draw out the quantitative relation between the shear wave velocity from the PS logging and the rock mass rating (RMR) determined from the inspection of the cores recovered from the same boreholes, These correlations were then used to relate the shear-wave velocity derived from REMI to RMR along the entire profile. The correlation between shear wave velocity and RMR was very good and so it was possible to estimate the RMR of the total zone of interest for the design of underground tunnel,

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.85.2-86
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• 2003

Fusarium graminearum is an important pathogen of cereal crops in many areas of the world causing head blight and ear rot of small grains. In addition to serious economic losses, this fungus produces mycotoxins, such as trichothecenes and zearalenone on diseased crops and has been a potential threat to human and animal health. To massively identify pathogenesis-related genes from F. graminearum, two representative strains (SCKO4 from rice and Z03643 from wheat) were mutagenized using restriction enzyme-mediated integration (REMI). In total, 20,DOD REMI transformants have been collected from the two strains. So far, 63 mutants for several traits involved in disease development such as virulence, mycotoxin production, and sporulation have been selected from 3,000 REMI transformants. Now, selected mutants of interest have being genetically analyzed using a newly developed outcross method (See Jungkwan Lee et al poster). In addition, cloning and characterization of genomic DNA regions flanking the insertional site in the genome of the mutants are in progress.

• The Plant Pathology Journal
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• v.22 no.3
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• pp.215-221
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• 2006

Gibberella zeae (anamorph: Fusarium graminearum) is an important pathogen of cereal crops. This fungus produces a broad range of secondary metabolites, including polyketides such as aurofusarin (a red pigment) and zearalenone (an estrogenic mycotoxin), which are important mycological characteristics of this species. A screen of G. zeae insertional mutants, generated using a restriction enzyme-mediated integration (REMI) procedure, led to the isolation of a mutant (Z43R606) that produced neither aurofusarin nor zearalenone yet showed normal female fertility and virulence on host plants. Outcrossing analysis confirmed that both the albino and zearalenone-deficient mutations are linked to the insertional vector in Z43R606. Molecular characterization of Z43R606 revealed a deletion of at least 220 kb of the genome at the vector insertion site, including the gene clusters required for the biosynthesis of aurofusarin and zearalenone, respectively. A re-creation of the insertional event of Z43R606 in the wild-type strain demonstrated that the 220-kb deletion is responsible for the phenotypic changes in Z43R606 and that a large region of genomic DNA can be efficiently deleted in G. zeae by double homologous recombination. The results showed that 52 putative genes located in the deleted genomic region are not essential for phenotypes other than the production of both aurofusarin and zearalenone. This is the first report of the molecular characterization of a large genomic deletion in G. zeae mediated by the REMI procedure.

• Journal of Microbiology
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• v.43 no.5
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• pp.417-423
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• 2005

In this study, the nematode-trapping fungus, Monacrosporium sphaeroides, was transformed with a plasmid harboring the hygromycin B phosphotransferase gene, via restriction enzyme-mediated integration (REMI). Frequencies of up to 94 transformants ${\mu}g^{-1}$ per linearized plasmid DNA were obtained by optimizing the PEG concentration, as well as the category and quantity of the added restriction enzyme. $90\%$ of the transformants were determined to be stable for drug resistance when 20 randomly selected transformants were tested. Southern analyses revealed that the transforming DNA was integrated into the M. sphaeroides genome either with or without rearrangement. Five mitotic stable mutant strains were obtained using this approach, all of which had been altered with regard to sporulation capacity and pathogenicity toward nematodes. Southern blot analyses of the five mutants revealed that foreign plasmid DNA had integrated into the genome. Three of the mutants, Tms2316, Tms3583 and Tms1536, exhibited integration at a single location, whereas the remaining two, Tms32 and Tms1913, manifested integration at double or multiple locations. Our results suggest that the transformation of M. sphaeroides via REMI will facilitate insertional mutagenesis, the functional analysis of a variety of genes, and the tagging or cloning of genes of interest.

• Proceedings of the Microbiological Society of Korea Conference
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• 2005.05a
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• pp.217.2-217
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• 2005

• Proceedings of the Microbiological Society of Korea Conference
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• 2005.05a
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• pp.224.3-224
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• 2005

• Journal of the Korean earth science society
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• v.33 no.7
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• pp.596-607
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• 2012

The R-Z relationship is one of important error factors to determine the accuracy of radar rainfall estimation. In this study, we have explored the effect of the R-Z relationships derived from disdrometer data in estimating the radar rainfall. The heavy rain event that produced flooding in St-Remi, Quebec, Canada has been occurred. We have tried to investigate the severity of rain for this event using high temporal (2.5 min) and spatial resolution ($1^{\circ}$ by 250 m) radar data obtained from the McGill S-band radar. Radar data revealed that the heavy rain cells pass directly over St-Remi while the coarse raingauge network was not sufficient to detect this rain event. The maximum 30 min (1 h) accumulation reaches about 39 (42) mm in St-Remi. During the rain event, the two disdrometers (POSS; Precipitation Occurrence Sensor System) were available: One used for the reflectivity calibration by comparing disdrometer Z and radar Z and the other for deriving disdrometric R-Z relationships. The result shows the significant improvement with the disdrometric reflectivity-dependent R-Z relationships against the climatological R-Z relationship. The bias in radar rain estimation is reduced from +12% to -2% and the root-mean squared error from 16 to 10% for daily accumulation. Using the estimated radar rainfall rate with disdrometric R-Z relationships, the flood event was well captured with proper timing and amount.