• 미생물학회지
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• 제23권4호
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• pp.309-314
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• 1985

Cellulose를 이용하는 Cellulomonas sp. CS 1-1을 육성하는 방법으로 원형질본합법을 응용하기 위해 영향요구변이주 간의 융합을 시도하였다 원형질본 형성 및 이들의 봉합 및 재생의 최적조건은 다음과 같다. 0.5M sucrose를 함유하는 복합배지에서 6시간 배양하고 0.2$\mu\textrm{m}$ penicillin G로 90분 처리한다. 이들 균체를 분리하여 0.2 mg/ml lysozyme으로 2시간 동안 처리하여 원형질체를 만들고, 이들을 40% polyethyleneglycol로 15분 처리하여 $10^{-3}$ ~$10^{-5}$의 빈도로 속운자 재조합이 이루어졌다. 원형질본의 재생에서 곰팡이의 오염을 방지하기 위해 Nystatin을 이용할 수 있었다.

• 대한의생명과학회지
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• 제28권2호
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• pp.83-91
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• 2022

This study was performed to investigate the contamination status of microorganisms that cause food poisoning in shellfish distributed in the metropolitan area of South Korea. The analyses were conducted according to the sample type, season, and region. In particular, the genotype of food poisoning viruses that explosively cause collective food poisoning was analyzed. Total 483 each of four types of shellfish (Crassostrea gigas, Mytilus coruscus, Pectea albicans albicans, and Scapharca subcrenata) were collected from four distribution sites located in the metropolitan area. We investigated foodborne pathogen by multiplex PCR and RT/Nested PCR from shellfish. As a result, Vibrio parahaemolyticus and Bacillus cereus were detected in 13 and 21 samples and Norovirus (NoV) GI and GII were detected in 7 and 12 samples, respectively. V. parahaemolyticus and NoV GI and GII showed differences in types of shellfish and seasons, and no correlation was confirmed with regional differences. Also, as a result of genotyping for the detected NoV GI and GII, they were confirmed as NoV GI.7, GI.5 and GII.3, which are causes of collective food poisoning. Therefore, cross-infection with shellfish can cause food poisoning. In particular, attention must be paid to the handling and cooking of shellfish in organizations that implement group feeding, and it is necessary to establish a management system for microorganisms that cause food poisoning in the process of shellfish farming and distribution.

• Parasites, Hosts and Diseases
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• 제60권3호
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• pp.195-200
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• 2022

There have been few reports on extra-enteric infections by Blastocystis STs and none have been molecularly identified in samples from human reproductive organs. We report for the first time the identification of 3 different subtypes of Blastocystis (ST1-3) in vaginal and sperm samples, from patients infected with Trichomonas vaginalis. Blastocystis STs were identified by PCR-sequencing and by phylogenetic inferences using 28 vaginal swab samples and 7 sperm samples from patients trichomoniasis. Blastocystis STs were identified in 6 of 28 vaginal swabs (21.4%) and in 3 of 7 sperm samples (42.8%). In both biological samples, STs 1-3 were found; one vaginal sample showed subtype co-infection with ST1 and ST3. High genetic variation was observed in the sequences obtained and no specific clustering in the phylogenetic trees was detected. Most of the haplotypes identified were placed far from the main dispersal centers. Our finding suggested that incorrect cleaning of the genital area or a contamination by combination of anal and vaginal intercourse.

• 한국미생물·생명공학회지
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• 제51권1호
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• pp.1-9
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• 2023

Arsenic (As), which is ubiquitous throughout the environment, represents a major environmental threat at higher concentration and poses a global public health concern in certain geographic areas. Most of the conventional arsenic remediation techniques that are currently in use have certain limitations. This situation necessitates a potential remediation strategy, and in this regard bioremediation technology is increasingly important. Being the oldest representativse of life on Earth, microbes have developed various strategies to cope with hostile environments containing different toxic metals or metalloids including As. Such conditions prompted the evolution of numerous genetic systems that have enabled many microbes to utilize this metalloid in their metabolic activities. Therefore, within a certain scope bacterial isolates could be helpful for sustainable management of As-contamination. Research interest in microbial As(III) oxidation has increased recently, as oxidation of As(III) to less hazardous As(V) is viewed as a strategy to ameliorate its adverse impact. In this review, the novelty of As(III) oxidation is highlighted and the implication of As(III)-oxidizing microbes in environmental management and their prospects are also discussed. Moreover, future exploitation of As(III)-oxidizing bacteria, as potential plant growth-promoting bacteria, may add agronomic importance to their widespread utilization in managing soil quality and yield output of major field crops, in addition to reducing As accumulation and toxicity in crops.

• 한국균학회지
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• 제50권3호
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• pp.225-233
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• 2022

Pyogo (Shiitake, Lentinula edodes) is one of the most important edible mushrooms because of its outstanding nutritive and medicinal value. In the registration and protection procedure for newly developed mushroom cultivars, the application of molecular markers that can supplement the morphological characteristic-based distinction has been strongly requested. Sanjo 701 and Chamaram, newly developed at the Federation Forest Mushroom Research Center of Korea, have been characterized as innovative cultivars suitable for customer demands because of their high yields and cultivation rates. However, no technical tools can protect the rights to these important cultivars. In this study, using comparative genomic information from 23 commercially available pyogo cultivars, we identified single nucleotide polymorphisms (SNPs) that accurately differentiated Sanjo701 and Chamaram from the other cultivars. We also developed high-resolution melting analysis (HRM)-based SNP markers that discriminate among the tested 23 pyogo cultivars. The developed SNP markers can be utilized for rapid, accurate identification of pyogo cultivars with low genetic diversity and to prevent cultivar contamination caused by illegally distributed inocula. In addition, these markers can serve as a crucial scientific basis for securing the right to conserve new cultivars in international markets.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2013년도 제45회 하계 정기학술대회 초록집
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• pp.88-89
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• 2013

A variety of influenza A viruses from animal hosts are continuously prevalent throughout the world which cause human epidemics resulting millions of human infections and enormous industrial and economic damages. Thus, early diagnosis of such pathogen is of paramount importance for biomedical examination and public healthcare screening. To approach this issue, here we propose a fully integrated Rotary genetic analysis system, called Rotary Genetic Analyzer, for on-site detection of influenza A viruses with high speed. The Rotary Genetic Analyzer is made up of four parts including a disposable microchip, a servo motor for precise and high rate spinning of the chip, thermal blocks for temperature control, and a miniaturized optical fluorescence detector as shown Fig. 1. A thermal block made from duralumin is integrated with a film heater at the bottom and a resistance temperature detector (RTD) in the middle. For the efficient performance of RT-PCR, three thermal blocks are placed on the Rotary stage and the temperature of each block is corresponded to the thermal cycling, namely $95^{\circ}C$ (denature), $58^{\circ}C$ (annealing), and $72^{\circ}C$ (extension). Rotary RT-PCR was performed to amplify the target gene which was monitored by an optical fluorescent detector above the extension block. A disposable microdevice (10 cm diameter) consists of a solid-phase extraction based sample pretreatment unit, bead chamber, and 4 ${\mu}L$ of the PCR chamber as shown Fig. 2. The microchip is fabricated using a patterned polycarbonate (PC) sheet with 1 mm thickness and a PC film with 130 ${\mu}m$ thickness, which layers are thermally bonded at $138^{\circ}C$ using acetone vapour. Silicatreated microglass beads with 150~212 ${\mu}L$ diameter are introduced into the sample pretreatment chambers and held in place by weir structure for construction of solid-phase extraction system. Fig. 3 shows strobed images of sequential loading of three samples. Three samples were loaded into the reservoir simultaneously (Fig. 3A), then the influenza A H3N2 viral RNA sample was loaded at 5000 RPM for 10 sec (Fig. 3B). Washing buffer was followed at 5000 RPM for 5 min (Fig. 3C), and angular frequency was decreased to 100 RPM for siphon priming of PCR cocktail to the channel as shown in Figure 3D. Finally the PCR cocktail was loaded to the bead chamber at 2000 RPM for 10 sec, and then RPM was increased up to 5000 RPM for 1 min to obtain the as much as PCR cocktail containing the RNA template (Fig. 3E). In this system, the wastes from RNA samples and washing buffer were transported to the waste chamber, which is fully filled to the chamber with precise optimization. Then, the PCR cocktail was able to transport to the PCR chamber. Fig. 3F shows the final image of the sample pretreatment. PCR cocktail containing RNA template is successfully isolated from waste. To detect the influenza A H3N2 virus, the purified RNA with PCR cocktail in the PCR chamber was amplified by using performed the RNA capture on the proposed microdevice. The fluorescence images were described in Figure 4A at the 0, 40 cycles. The fluorescence signal (40 cycle) was drastically increased confirming the influenza A H3N2 virus. The real-time profiles were successfully obtained using the optical fluorescence detector as shown in Figure 4B. The Rotary PCR and off-chip PCR were compared with same amount of influenza A H3N2 virus. The Ct value of Rotary PCR was smaller than the off-chip PCR without contamination. The whole process of the sample pretreatment and RT-PCR could be accomplished in 30 min on the fully integrated Rotary Genetic Analyzer system. We have demonstrated a fully integrated and portable Rotary Genetic Analyzer for detection of the gene expression of influenza A virus, which has 'Sample-in-answer-out' capability including sample pretreatment, rotary amplification, and optical detection. Target gene amplification was real-time monitored using the integrated Rotary Genetic Analyzer system.

• 한국축산식품학회지
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• 제29권6호
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• pp.695-701
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• 2009

본 연구는 축산물가공장 작업환경에 설정된 CCP의 유효성을 평가하기 위하여, 원료도체에서부터 포장에 이르는 전 공정에서 Escherichia coli를 검출하고 PFGE genotyping으로 genetic strain의 흐름을 분석하여 CCP의 적합성을 평가하였다. 시료채취를 위한 작업장은 도축장에서 냉장차량으로 원료도체를 공급받는 독립형 소규모 HACCP 가공장(100두 처리/일)과 축산물종합처리장내 HACCP 가공장(500두 처리/일)을 각각 1곳씩 선정하였다. 시료는 원료도체, 시설 및 장비(발골기, 박피기, 작업대, 벨트, 칼, 면장갑), 환경(벽, 바닥)과 작업공정상의 발골육, 부분육, 최종육을 무균적으로 채취하였다. 작업공정에서 E. coli의 높은 분리율(59.6%)과 작업환경 전반에서의 고른 검출로 E. coli가 축산물가공장 위생지표균으로의 활용 가능성을 확인할 수 있었다. 분리된 E. coli의 genotype 분포는 작업장내 미생물 오염의 흐름을 보여주었다. 소규모가공장의 경우 오염된 원료육 반입이 작업공정을 오염시키고 공정중의 교차오염 발생으로 최종육에 영향을 주었고, 축산물종합처리장내 작업장은 원료육 오염보다는 작업환경에 상존하는 오염미생물의 교차오염이 최종육에 영향을 주는 양상이었다. 따라서 포장육가공장과 같이 작업공정에 멸균처리 과정이 없는 경우는 선행요건에 해당되는 원료반입과 작업환경의 세척 및 소독이 CCP로 관리되거나 이에 준하는 특별관리가 필요하였다. 결과적으로, 작업공정에서의 E. coli 검출과 genotyping은 작업공정상의 미생물 오염의 흐름을 파악하는데 유용하였으며, 이러한 방법은 HACCP 작업장의 미생물 오염원 분석과 설정된 생물학적 critical control point의 적합성 평가에 효과적이었다.

• 자원환경지질
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• 제37권1호
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• pp.1-19
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• 2004

국내 금속광상의 성인적 유형은 주로 열수충진형 광상, 스카른형 광상, 열수교대형 광상 및 변성퇴적형 광상으로 배태되고 이외에 일부는 정마그마형 광상, 반암형 광상, 알라스카이트형 광상 등으로 부존되고 있다. 이러한 폐광산으로부터 발생되는 수질 및 토양의 환경문제는 개발규모 및 수반금속, 주변암상과 관련된 부존특성과 함께 열수변질작용과 관련된 광물조합, 광석-맥석 광물의 조직, 유형 및 양적관계, 풍화의 진행단계와 관련된 이차-삼차광물의 존재형태 등의 광물-지화학적 특성에 좌우되고 있다. 1970년대 초반부터 1990년대 후반에 걸쳐 광산개발이 종료된 국내금속광상은 폐광 이후의 경과기간에 따라 황화-황염광물과 같은 일차광물로부터 용해되어 수산화광물, 황산염광물, 탄산염광물로 변화하는 과정에서 산화-용출-침전-재용출-이동에 의한 다단계의 복잡한 지화학적 반응관계를 보이고 있다. 금속광상의 산성 배수 및 중금속오염은 다금속 광화작용과 관련된 비철금속 광상과 동시에 맥상광상 및 각력파이프형 광상과 백악기금광상에서 가장 높은 용출가능성을 보이는 반면, 스카른형 광상, 열수교대형 광상, 정마그마형 광상, 맥상 금-은광상, 맥상 동광상 및 변성퇴적형 광상에서 오염가능성은 매우 낮은 경향을 보이고 있다. 이와 같이 광상 유형과 관련된 지질부존 특성은 폐석 및 광미를 구성하는 황화광물 및 탄산염광물의 유형 및 양비와 밀접한 연관성을 갖고 있으며, 산성 배수에 함유된 금속원소종의 유형 및 용출량과 같은 지화학적 특성에 직접적으로 영향을 미치고 있다.

• 한국버섯학회지
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• 제15권4호
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• pp.273-278
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• 2017

느티만가닥버섯은 맛과 기능성이 풍부한 버섯으로 많이 활용되고 있다. 하지만 긴 재배기간과 낮은 자실체 수확량, 균이 오랜시간 배양되어 오염이 쉽게 발생되는 문제점을 극복할 새로운 품종육성이 필요하다. 이에 따라 육종모본으로 활용되는 느티만가닥 55균주의 종내 유전자원의 정확한 정보를 얻고자 분자유전학적 ISSR 마커분석을 활용하였다. 사용된 마커 중에서 ISSR 13과 15 마커를 사용했을 때 다형성이 분석되었으며 특히 ISSR 15마커 분석으로 다형성이 쉽게 구분되었다. UPGMA분석법으로 계통도를 분석하였을 때, 일부 백색을 가지고 있는 KMC03106, KMC03107, KMC03108 3 균주가 두 마커 모두에서 가까운 유연관계를 가졌으며, ISSR 15는 수집년도에 따라 3개의 그룹으로 구분되는 것을 확인할 수 있었다. 이 결과로 ISSR마커의 다형석 분석은 느티만가닥버섯의 몇몇 균주에서는 갓색의 유연관계 확인과 수집 시기에 따른 유전변이 구분이 가능하며 자원의 유전적 다양성 확인할 수 있어, 느티만가닥버섯의 품종육성을 위한 효율적인 모본 선발이 가능 할 것으로 사료된다.

• 한국환경보건학회지
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• 제39권2호
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• pp.166-177
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• 2013

Objectives: The genus Legionella is common in aquatic environments. Some species of Legionella are recognized as potential opportunistic pathogens for human, notably Legionella pneumophila that causes, Legionellosis. Thus, we investigated the contamination of Legionella pneumophila on water supply systems in Seoul, including cooling towers, public baths, hospitals and fountains. Methods: The existence of 16S rRNA and mip gene of L. pneumophila was confirmed in the genome of the isolated strains by PCR. Results: During the summer season of 2010 and 2011, Legionella pneumophila were detected from 163 samples (21.1%) out of 772 samples collected. Among the 163 strains of L. pneumophila, eighty one isolates belonged to serogroup 1 (57.4%), 23 isolates were serogroup 5 (16.3%), 21 isolates were serogroup 6 (14.9%), 8 isolates were serogroup 2 (5.79%), and 8 isolates were identified in serogroup 3 (5.7%). Through PFGE (pulsed-field gel electrophoresis) analysis using Sfi I, genetic types of L. pneumophila were classified into five (A to E) patterns by the band similarity with excess of 70% from public baths. Conclusions: The PFGE patterns of the serotypes showed a tendency for diversity of L. pneumophila. Our results suggest the existence of serological and genetic diversity among the L. pneumophila isolates.