• Korean Journal of Microbiology
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• v.37 no.2
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• pp.137-144
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• 2001

In order to investigate the diversity of bacterial community in the mud flat of Sunchon Bay, Chunnam province, diversity of amplified 16S rDNA was examined. Total DNA was extracted from sediment soils and 16S rDNAs were amplified using PCR primers based on the universally conserved sequences in bacteria. Clonal libraries were constructed and 111 clones were examined by amplified rDNA restriction analysis (ARDRA) using HaeIII. Clones were clustered based on restriction patterns using computer program, GelCompar II. One hundred different RFLP types were detected from 111 clones. The 20 clones were selected and sequenced according to dendrograms derived from ARDRA, to cover most of the bacterial diversity in the clone libraries. None of the clones were identical to any representatives in the Ribosomal Database Project small subunit RNA databases and GenBank. All sequences showed between 77 and 96.8% similarity to the known 16s rRNA sequence from cultured organisms. The 20 clones sequenced fell into seven major lineages of the domain Bacteria: alpha-, delta-, gamma-Proteobacteria, low G+C Gram positive bacteria, high G+C Gram positive bacteria, Sphingobacteria (Cytophaga) and Cyanobacteria (chloroplast). Among the clones, the Proteobacteria were dominant.

• The Korean Journal of Mycology
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• v.31 no.2
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• pp.103-113
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• 2003

Twenty five isolates of Alternaria were obtained from various solanaceous crops in Korea. For all isolates, morphological characteristics of the conidia were determined and compared with those of representative isolates of A. solani and A. tomatophila. A selection of the isolates and the representative Alternaria isolates were evaluated for Pathogenicity to potato, tomato, egg plant and red pepper. Molecular characteristics of 17 isolates of Alternaria inculding the representative isolates were determined using sequence analysis of IRS rDNA and histone H3 gene, and URP-PCR analysis. Based on morphological characteristics, the isolates from the solanaceous crops were grouped as identical or very similar to either A. tomatophila (ATO), A. solani (ASO), and unidentified Alternaria sp. (ASP). Isolates of ASO were moderately pathogenic to all the solanaceous crops tested, but ATO isolates were highly pathogenic to tomato and the ASP isolate was pathogenic only to potato. Among the molecular markers used in this study, the URP-PCR analysis was found to be appropriate for taxonomic resolution of these species. Based on the conidial morphology, pathogenicity test and molecular characteristics, A. tomatophila (early blight of tomato) could be distinguished from A. solani (early blight of potato), and the Alternaria sp. (ASP) from potato, which was closely related to ASO in conidial morphology, was considered as a new species.

• Tuberculosis and Respiratory Diseases
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• v.44 no.2
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• pp.251-263
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• 1997

Background : The emergence of multidrug-resistant strains of Mycobacterium tuberculosis presents a significant challange to the treatment and control of tuberculosis, and there is an urgent need to understand the mechanisms by which strains acquire multidrug resistance. Recent advances in molecular methods for the detection of M. tuberculosis genetic targets have approached the sensitivity of culture. Furthermore the prospect of determining resistance in mycobacteria at the nucleic acid level particulary to first-line drugs like rifampin, isoniazid has provided a glimps of the next generation of sensitivity test for M. tuberculosis. Previous studies in RMP resistant M. tuberculosis have shown that mutation in $\beta$subunit of RNA polymerase is main mechanism of resistance. Method : In this study, rpoB gene for the $\beta$subunit of RNA polymerase from M. tuberculosis of 42 cultured samples (32 were RMP resistant and 10 were sensitive cases) were isolated and characterised the mutations. Direct sequencing data were compared with the results of INNO-LiPA Line Probe Assay (LiPA, Innogenetics, Belgium), commercial RMP resistance detecting kit using reverse hybridization method. Results : All of the RMP resistant samples were revealed the presence of mutation by LiPA. In 22 samples (68.8%) out of 32 RMP resistant cases, the mutation types were confirmed by the positive signal at one of 4 mutation bands in the strip. The most frequent type was R5 (S531L) which were 17 cases (77.3%). Results of direct sequencing were identified the exact characteristics of 8 mutations which were not confirmed by LiPA. S522W type point mutation and 9 base pair deletion at codon 513~515 were new identified mutations for the first time. Conclusion : Mutations in rpoB gene is the main mechanism of RMP resistance in M. tuberculosis and LiPA is a very useful diagnostic tool for the early diagnosis of RMP resistance in M. tuberculosis.

• Development and Reproduction
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• v.5 no.2
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• pp.107-114
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• 2001

The recent reports that endocrine disruptors(EDs) bring about abnormalities in reproductive organs and functions of invertebrates suggest that mammals be affected by the EDs. The present study examined the influence of 2-bromopropane(2-BP) by looking at the sexes of litters in mouse. The expression of sex-related genes during sex differentiation was also investigated in the fetus of mouse. The male and female mice were infused with 2-BP for 3 weeks before mating. The litters were sexed at the weaning time from the 4 different groups. The sex-related genes were identified by RT-PCR from the fetuses at gestation 10 days. The sequences of the genes were analysed by comparing to those of other animals. The mean numbers of litters survived by the weaning time were slightly reduced in the only group of both female and male mice treated with 2-BP. The female litters were greater than male litters in the only group of female treated with 2-BP. The other groups showed male litters greater than female litters. The sex-related genes, SRY, DAX1, SF1 , and AMH genes were identified and sequenced, showing 416, 466, 326, 389 base pairs, respectively. All of the genes had the homology of 89～90％ with rat and 81～92％ with human within the range of bases identified. They were expressed at the time of sex determination. Therefore, it appears that 2-BP somewhat affects the reproductive activity of adult mouse. Influence of 2-BP on the reproductive function is expected to be studied through the expression of the sex-related genes.

• The Korean Journal of Mycology
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• v.39 no.1
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• pp.22-30
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• 2011

Weonhyeong is one of important commercial strains. It has good characteristics of bundle formation, grey colored pilei and high productivity. We previously reported grouping of 70 strains of Pleurotus ostreatus in which one group contained 35 strains including Weonhyeong. Four strains in that group showed same profiles implicating no variety distinction for mushroom cultivation. Now we developed a specific marker for identification of Weonhyeong. Sequence Characterized Amplified Region (SCAR) marker was developed from the RAPD amplicon. SCAR marker 'S-OPO5' produced only one band specific to 2183, 2240, 2595 and 2725 strains showing similar banding patterns to Weonhyeong in RAPD-PCR results. The sequence of 'S-OPO5' marker was unknown when compared with the data in the Genbank using BLASTN. BLASTX results indicated that the marker showed significant alignment with the protein sequences in Tricholoma bakamatsutake reverse transcriptase. The results indicate that this new SCAR marker ('S-OPO5') will be valuable to distinguish the Weonhyeong similar strains from Pleurotus spp.

• The Korean Journal of Blood Transfusion
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• v.29 no.3
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• pp.310-319
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• 2018

Background: Research on next-generation sequencing (NGS)-based HLA typing is active. To resolve the phase ambiguity and long turn-around-time of conventional high resolution HLA typing, this study developed a NGS-based high resolution HLA typing method that can handle large-scale samples within an efficient testing time. Methods: For HLA NGS, the condition of nucleic acid extraction, library construction, PCR mechanism, and HLA typing with bioinformatics were developed. To confirm the accuracy of the NGS-based HLA typing method, the results of 192 samples HLA typed by SSOP and 28 samples typed by SBT compared to NGS-based HLA-A, -B and -DR typing. Results: DNA library construction through two-step PCR, NGS sequencing with MiSeq (Illumina Inc., San Diego, USA), and the data analysis platform were established. NGS-based HLA typing results were compatible with known HLA types from 220 blood samples. Conclusion: The NSG-based HLA typing method could handle large volume samples with high-throughput. Therefore, it would be useful for HLA typing of bone marrow donation volunteers.

• The Korean Journal of Mycology
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• v.39 no.1
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• pp.31-38
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• 2011

In this study, 81 commercial strains of Pleurotus species cultivated in South Korea were analyzed with randomly amplified polymorphic DNA (RAPD) technique. Sequence characterized amplified region (SCAR) markers were developed by designing from one RAPD polymorhic band specific to Suhan strain. The SCAR primer pair 'S-OPA13-1' amplified a 590-bp fragment in the varieties originated from Suhan strain. The Blast search of S-OPA13-1 showed high homology to the POMFBO1 P. ostreatus cDNA clone MFB02-A05 and Laccaria bicolor S238N-H82. The results showed that this SCAR marker can clearly distinguish Suhan strains from Pleurotus spp.

• Korean Journal of Ecology and Environment
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• v.38 no.2 s.112
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• pp.137-145
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• 2005

Traditional screening techniques have missed up to 99% of microbial resources existing in the nature. Strategies of direct cloning of environmental DNAs comprising tine genetic blueprints of entire microbial metagenomes provide vastly more genetic information than is contained in the culturable. Therefore, one way to screening the useful gene in a variety of environments is the construction of metagenomic DNA library. In this study, the water samples were collected from Daecheong Reservoir in the mid Korea, and analyzed by T-RFLP to examine the diversity of the microbial communities. The crude DNAs were extracted by SDS-based freezing-thawing method and then further purified using an $UltraClean^{TM}kit$ (MoBio, USA). The metagenomic libraries were constructed with the DNAs partially digested with EcoRI, BamHI, and SacII in Escherichia coli DH10B using the pBACe3.6 vector. About 14.0 Mb of metagenomic libraries were obtained with average inserts 13 ${\sim}$ 15 kb in size. The genes responsible for degradation of 1, 2-dichloroethane (1, 2-DCE) via hydrolytic dehalogenation were identified from the metagenomic libraries by colony hybridization. The 1, 2-dichloroethane dehalogenase gene (dhlA) was cloned and its nucleotide sequence was analyzed. The activity of the 1, 2-DCE dehalogenase was highly expressed to the substrate. These results indicated that the dhlA gene identified from the metagenomes derived from Deacheong Reservoir might be useful to develop a potent strain for degradation of 1, 2-DCE.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.19 no.3
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• pp.382-388
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• 2018

In this study, the SELEX method was used to screen for and select aptamers with high selectivity and affinity for Streptococcus mutans, which is the causative agent of dental caries. Aptamers are single stranded oligonucleotides of DNA or RNA with high selectivity and affinity for target molecules because of their specific three-dimensional structures that are used to collect biometric information. When compared to antibodies in vitro, aptamers are more advantageous because of their ease of use in the screening process, low cost, chemical stability, and lack of restrictions on the target molecules. We sorted DNA aptamers, which contain 44 bp or 38 bp primer sequences in 5' and 3', 39 bp random sequences in the middle.We then analyzed the character and affinity to S. mutans. Aptamers of specific oligonucleotides that combine with S. mutans were selected and these results are selectively fused to S. mutans. The results confirmed that DNA aptamers can be used for rapid diagnosis and treatment of dental caries caused by bacteria of the oral cavity, namely S. mutans.

• Journal of Science and Technology Studies
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• v.17 no.1
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• pp.71-115
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• 2017

In 1953, theoretical physicist George Gamow attempted to explain the process of protein synthesis by hypothesizing that the base sequence of DNA encodes a protein's amino acid sequence and, in response, proposed the nucleic acid-protein information transfer model, which he dubbed the "diamond code." After expressing interest in discussing the daring hypothesis, contemporary biologists, including James Watson, Francis Crick, Sydney Brenner, and Gunther Stent, were soon invited to join the RNA Tie Club, an informal research group that would also count biologists and various researchers in physics, mathematics, and computer engineering among its members. In examining the club's formation, growth, and decline in multidisciplinary research on deciphering the genetic code in the 1950s, this paper first investigates whether Gamow's idiosyncratic approach could be adopted as a collaborative research forum among contemporary biologists. Second, it explores how the RNA Tie Club's research agenda could have been expanded to other relevant research topics needing multidisciplinary approach? Third, it asks why and how the RNA Tie Club dissolved in the late 1950s. In answering those questions, this paper shows that analyses on the intersymbol correlation of the overlapping code functioned to integrate diverse approaches, including sequence decoding and statistical analysis, in research on the genetic code. As those analyses reveal, the peculiar approaches of the RNA Tie Club could be regarded as a useful method for biological research. The paper also concludes that the RNA Tie Club dissolved in the late 1950s due to the disappearance of the collaborative research agenda when the overlapping code hypothesis was abandoned.