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RT-PCR과 nested PCR을 이용한 Nepovirus속 식물검역 바이러스 4종의 정밀진단
이시원,강은하,신용길,이수헌 한국식물병리학회 2013 식물병연구 Vol.19 No.3
For quarantine purpose, we developed the RT- and nested PCR module of Tomato black ring virus (TBRV),Arabis mosaic virus (ArMV), Cherry leafroll virus (CLRV) and Grapevine fanleaf virus (GFLV). The PCRmodules, developed in this study make diagnosis more convenient and speedy because of same PCRcondition. And also, the methods are more accurate because it can check whether the result is contaminationor not using the mutation-positive control. We discard or return the 27 cases of Nepovirus infection seed byemploying the module past 3 years. This study provides a rapid and useful method for detection of fourquarantine plant viruses. 본 연구에서는 식물검역바이러스 4종(TBRV, ArMV,CLRV 및 GFLV)을 RT-PCR과 nested PCR 방법으로 진단 할 수 있는 방법을 개발하였다. 본 연구에서 개발한방법은 모두 같은 PCR 조건으로 검사자에게 편리성과 신속성을 높여줄 뿐 아니라, 돌연변이-양성대조구의 사용으로 실험 오염여부를 확인할 수 있어 더욱 정확하다. 개발한 방법으로 최근 3년 Nepovirus속 4종의 바이러스를 검사한 결과, 27건을 검출하여 검역처분 하였다. 본 연구 결과들은 앞으로도 수출입 식물에서 해당 바이러스들을 신속, 정밀하게 진단할 수 있는 방법으로 활용할 수 있을것으로 기대된다.
이시원,정준금,민병익 한국정책분석평가학회 2011 政策分析評價學會報 Vol.21 No.3
본 연구에서는 정책결정이 다양한 이해관계자가 참여한 가운데 동태적으로 이루어진다는 것을 전제로 특정 정책의 결정에 지지집단과 저항집단의 존재여부 그리고 지지와 저항의 강도가 실제 정책결정의 소요시간에 어떠한 영향을 미치고 있는지를 살펴보고자, 우리나라 중앙정부의 주요 정책 219개를 실증적으로 분석하였다.본 연구의 분석결과에 의하면, 정책결정에 참여하는 참여자의 특성을 지지적인 태도를 가지고 있는 지지집단과 저항적인 태도를 보이고 있는 저항집단, 그리고 행정부 내부의 관련기관의 호의적인 태도정도로 구분하여 살펴 본 후, 이들의 결합관계에 따라 정책결정 소요시간이 어떻게 달라지는가를 아울러 파악하였다. 이들 결합관계와 정책결정 소요시간의 관계에서 확인된 결과는 지지집단의 여부는 참여자의 수를 늘리는 정도의 효과가 있을 뿐, 지지집단이 있다고 해서 정책결정 소요시간이 짧아지지는 않는다는 것이다. 정책결정소요시간에 중요한 영향을 미치는 요인은 저항집단의 여부와 저항의 강도, 그리고 행정부 내부 관련기관의 협력 정도라는 것을 확인할 수 있었다.
이시원,장준형,배경선,이원석,정현미,박상정 한국미생물학회 2018 The journal of microbiology Vol.56 No.12
This study investigated the occurrence of human Norovirus (HuNoV) by genotype in 1,486 groundwater samples collected from 843 groundwater wells suspected of contamination during 2007–2016, in South Korea. We identified and genotyped 186 HuNoV sequences in 178 HuNoV-positive samples using the RIVM-NoroNet norovirus genotyping tool (NGT) and phylogenetic tree analysis based on RIVM-NoroNet reference sequences. HuNoV GII was more prevalent than GI. The major genotypes detected were HuNoV GII.4 (43.0%), GII.22 (15.6%), GI.5 (10.2%), and GI.1 (8.6%); several genotypes accounted for < 5.0% of all HuNoVs, including GII.17, GI.6, GI.4, GII.6, GI.8, GII.3, GII.13, GI.3, GI.7, GI.2, GI.9, GII.1, GII.8, and GII.10. The prevalence of HuNoVs and number of genotypes detected has drastically decreased over the last decade. HuNoV GII.17, the emerging genotype worldwide including Europe and Asia, appeared in Korean groundwater from 2010, dominated in 2013–2014, and continued to be observed. HuNoV GII.4, the major type occurred last decade from Korean groundwater except 2013–2014, continued to be detected and prevalent similar to HuNoV GII.17 in 2016.
이시원,배경선,박지현,김진호,이진영,최지원,박응로,유경아 대한의생명과학회 2021 Biomedical Science Letters Vol.27 No.4
Human Astrovirus (HuAstV) is an important gastrointestinal pathogen that is frequently reported worldwide. Monitoring of contaminated groundwater has been suggested since HuAstV is transmitted through the fecal-oral route. This study developed a test method based on conventional reverse transcription (RT)-nested polymerase chain reaction (PCR) that involves SL® non-specific reaction inhibitor for unknown non-specific amplification taking place in the groundwater environment. An optimal method for detecting HuAstV in groundwater sample through analysis and comparison against conventionally reported method was also suggested. The developed method enabled the production of nested PCR amplicon of 630 nt, which is a sufficient length for similarity analysis based on sequencing and genotyping. Amplicons suspected to be HuAstV were amplified in two out of the twenty groundwater samples collected in Korea, presenting 99.77% and 99.73% similarity against HuAstV 1 strain lhar/2011/kor (JN887820.1) in sequencing, respectively. These amplicons were identified as HuAstV 1.
Methylobacterium dankookense sp. nov., Isolated from Drinking Water
이시원,오현우,Kang-Hyun Lee,안태영 한국미생물학회 2009 The journal of microbiology Vol.47 No.6
A pink-pigmented bacterium, designated SW08-7T was isolated from the drinking water of a water purifier. Cells were Gram-negative, rod-shaped, strictly aerobic, and non-spore-forming. It grew optimally at 25°C, pH 6~7. Phylogenetic analysis based on 16S rRNA gene sequence showed that strain SW08-7T belongs to the genus Methylobacterium. The highest 16S rRNA gene sequence similarities were found to Methylobacterium mesophilicum JCM 2829T (96.9%), Methylobacterium brachiatum B0021T (96.9%), Methylobacterium phyllosphaerae CBMB27T (96.6%), Methylobacterium radiotolerans JCM 2831T (96.6%), and Methylobacterium hispanicum GP34T (96.5%). DNA-DNA hybridization experiment revealed low-level (28.5%) of DNA- DNA relatedness between strain SW08-7T and Methylobacterium hispanicum. The genomic DNA G+C content was 68.9 mol% and the major isoprenoid quinone was Q-10. The major cellular fatty acid of strain SW08-7T was C18:1 ω7c (79.8±2.1%). Results of phylogenetic, phenotypic, and biochemical analyses revealed that strain SW08-7T could be classified as representing a novel species of genus Methylobacterium, for which the name Methylobacterium dankookense sp. nov. is proposed. The type strain is SW08-7T (=KCTC 22512T =DSM 22415T).