• 한국양식학회:학술대회논문집
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• 한국양식학회 2003년도 추계학술발표대회 논문요약집
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• pp.134-135
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• 2003

The geographic expansion of the toxic dinoflagellates genus Alexandrium has been shown to be world wide ranging. The members of the genus Alexandrium ocnstituted of 20-30 species did not show substantial differences in their morphology, which is mostly referred in the 'tamarensis species complex', except some species. Though rDNA sequences variations are very few and pseudogene types are so diverse that it is difficult to use them as the specific markers. In this study, we outlined Korean and Japanese A, tamarense and A. catenella regional isolates by phylogenetic analysis inferred from no cutting alignments of LSU rDNA D1-D2 and SSU rDNA sequences to group these regional isolates. The results were compared to RFLP patterns of PCR products targeted chloroplast DNA. Lastly screening of highly repeated microsatellite DNA which is frequently used for population analysis in eukaryotes was conducted. A. catenella regional strains identified by the sequencing of rDNA D1-D2 domain were divided into at least 3 groups of type E, CMC and Chinese type, divergence root may not be deep comparing with that of A. tamarense whose pseudogenes are very variable. Results of RFLP pattern and the phylogeny of the unknown gene targeting chloroplast showed that Korean and Japanese A. catenella regional isolates were divided into 3 types: Korean, Japanese and the third CMC types. Population-specific PCR amplification with Japanese A. catenella type-specific PCR primers was useful method for population analysis of A. catenella. Various types of satellite sequences such as 5 nucleotides repeats were obtained from A. tamarense and A. catenella. The 5 nucleotides repeats were primed at the both 3'and 5' ends, and these repeats were prominent as longer repeated motifs. This repeated DNA was intercalated as internal sequences containing various types subrepeats. It is expected that these satellite DNA would be a useful molecular population marker through detail comparison among Alexandrium regional isolates to trace their transferring pathway and to prevent their human-associated their regional extents.

• Journal of Ginseng Research
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• 제34권1호
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• pp.41-46
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• 2010

Expensive herbs such as ginseng are always a possible target for fraudulent labeling. New mountain ginseng strains have occasionally been found deep within mountain areas and commercially traded at exorbitant prices. However, until now, no scientific basis has existed to distinguish such ginseng from commonly cultivated ginseng species other than by virtue of being found within deep mountain areas. Polymerase chain reaction (PCR) analysis of the internal transcribed spacer has been shown to be an appropriate method for the identification of the most popular species (Panax ginseng) in the Panax ginseng genus. A single nucleotide polymorphism (SNP) has been identified between three newly found mountain ginseng (KGD4, KGD5, and KW1) and already established Panax species. Specific PCR primers were designed from this SNP site within the sequence data and used to detect the mountain ginseng strains via multiplex PCR. The established multiplex-PCR method for the simultaneous detection of newly found mountain ginseng strains, Korean ginseng, and foreign ginseng in a single reaction was determined to be effective. This study is the first report of scientific discrimination of "mountain ginsengs" and describes an effective method of identification for fraud prevention and for uncovering the possible presence of other, cheaper ginseng species on the market.

• Journal of Microbiology and Biotechnology
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• 제23권4호
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• pp.555-559
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• 2013

Vibrio cholerae O1 and O139 are the major serotypes associated with illness, and some V. cholera non-O1 and non-O139 isolates produce cholera toxin. The present study describes a quantitative polymerase chain reaction (qPCR) assay for the species-specific detection and quantitation of V. cholera using a primer pair based on an outer membrane lipoprotein lolB gene for the amplification of a 195 bp DNA fragment. The qPCR primer set for the accurate diagnosis of V. cholera was developed from publically available genome sequences. This quantitative PCR-based method will potentially simplify and facilitate the diagnosis of this pathogen and guide disease management.

• 한국균학회지
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• 제36권2호
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• pp.138-143
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• 2008

토마토 잿빛곰팡이병균(B. cinerea)은 비닐하우스에서 재배할 때 토마토의 꽃과 줄기의 감염을 통해 매우 심각한 피해를 입힌다. 이 연구에서 토마토에 발병하는 잿빛 곰팡이병균의 검출 및 종 동정을 위해 새로운 종 특이적 primer set가 개발되었다. 종 특이적 primer(BTF1/BTR1)는 B. cinerea와 유전적으로 매우 유사한 진균들의 pyruvate carboxylase(pyc) 유전자 내부의 변이영역으로부터 설계되었다. 10개의 다른 기주식물에서 분리된 13균주의 모든 B. cinerea에서 112 bp 크기의 PCR 산물들이 만들어졌다. 그러나 6종의 다른 Botrytis 속균, 4종의 Botryotinia 속균, 5종의 Sclerotinia 속균 및 그 이외 16속의 다른 식물병원균들에 대해서는 PCR 반응이 나타나지 않았다. 종 특이적 primer의 반응민감도 한계는 대략 2 pg이었다. 자연상태에서 B. cinerea에 감염된 토마토 식물체와 인공적으로 접종된 식물체로부터 종 특이적 primer를 활용한 병원균의 PCR 검출이 이루어졌다. 이 연구결과로 미루어 새롭게 개발된 primer는 높은 반응민감도와 종 특이성을 나타내 추후 토마토 잿빛곰팡이병의 빠른 진단 및 병원균의 정확한 동정에 활용될 수 있을 것으로 판단된다.

• Journal of Applied Biological Chemistry
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• 제58권4호
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• pp.383-387
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• 2015

국내 유통 식품 수입 식품 중 토끼와 고양이 고기의 혼입 여부를 알아내고 불법 도축된 고양이 고기를 토끼 고기나 다른 고기로 속여 판매하는 것을 방지하기 위해 토끼와 고양이를 동시에 검출할 수 있는 polymerase chain reaction (PCR) 법을 개발하였다. 토끼와 고양이의 종 특이 프라이머는 미토콘드리아의 cytochrome b 유전자를 대상으로 하였고 개발된 프라이머를 가공식품에 활용하는 것을 고려하여 PCR 산물의 크기는 토끼 101 bp, 고양이 191 bp로 최소화 하였다. 프라이머의 특이성은 총 21종의 동물을 대상으로 검토하였다. 개발된 검출법의 검출 한계는 시료 DNA를 희석하여 PCR과 Bioanalyzer로 확인한 결과 토끼는 0.005 ng, 고양이는 0.0005 ng이었다.

• 한국미생물·생명공학회지
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• 제36권2호
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• pp.96-100
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• 2008

recA 유전자를 특이적으로 증폭하는 PCR을 이용하여 우리나라 전통 침채류 발효에 관여하는 유산균의 다양성을 검토해 보았다. 김치에서 많이 검출되는 유산균 7종 및 대조군으로 Lactobacillus acidophilus를 검출할 수 있는 특이적 PCR primer pair을 이용하여 전통 침채류 5증(갓김치, 동치미, 배추김치, 오이소박이, 총각김치로부터 추출한 DNA를 template로 PCR을 수행한 결과, 5종의 침채류 모두에서 Lactobacillus plantarum과 Lactobacillus sakei가 검출되었지만, Lactobacillus paraplantarum, Lactobacillus pentosus와 대조군인 Lb. acidophilus는 검출되지 않았다. Lactobacillus brevis와 Leuconostoc citreum은 배추김치에서만 검출되었으며, Leuconostoc mesenteroides의 경우 갓김치, 동치미, 배추김치, 오이소박이에서 검출되었다. 주재료의 종류에 따라서 발효에 관여하는 유산균은 차이가 있는 것으로 추정되며, Lb. plantarum과 Lb. sakei가 우리나라 침채류 발효에 가장 널리 관여하는 것으로 사료된다.

• 대한수의학회지
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• 제55권2호
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• pp.105-110
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• 2015

A real-time PCR assay using hybridization probe (HybProbe) has been developed to detect Brucella (B.) melitensis strains. The primer and HybProbe sets were designed based on the gap gene of chromosome I with a specific single nucleotide polymorphism of B. melitensis. Specificity of the assay was confirmed by comparison to reference Brucella species and other related strains. In the melting curve analysis, B. melitensis generated a peak at $67^{\circ}C$ unlike those for other Brucella species observed at $61^{\circ}C$. Sensitivity of the assay for B. melitensis ranged from 20 ng to 200 fg of genomic DNA. The ability to identify 94 Mongolian B. melitensis isolates using the real-time PCR assay was identical to that of classical biotyping methods and differential multiplex PCR. These data showed that this new molecular technique is a simple and quick method for detecting B. melitensis, which will be important for the control and prevention of brucellosis.

• 한국산림과학회지
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• 제104권2호
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• pp.332-335
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• 2015

자낭균인 Taphrina wiesneri는 한국의 공원과 도로 주변에 주로 식재되는 왕벚나무에 빗자루병을 발생시킨다. 이 병원균은 생태적 특성이 잘 알려져 있지 않아 방제법 등을 개발하는데 어려움이 있다. 이번 연구에서는 빗자루 병원균의 월동부위를 조사하기 위해 빗자루 병징을 보이는 왕벚나무의 건전가지와 이병가지, 그리고 병징이 나타나지 않은 건전한 왕벚나무 가지에서 종 특이적 primer (TwITSF와 TwITSR)를 사용하여 월동부위를 조사하였다. 그 결과 빗자루병에 감염된 왕벚나무의 이병가지 뿐만 아니라 건전가지에서도 종 특이적인 PCR 증폭산물이 관찰되었으며, 그 이외의 건전한 왕벚나무와 감염목 주변의 다른 식물 종 샘플에서는 PCR 증폭산물이 관찰되지 않았다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVI)
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• pp.396-397
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• 2005

The polyene antibiotics are a family of most promising antifungal polyketide compounds, typically produced by actinomycetes species. Using the polyene CYP-specific PCR screening with served actinomycetes genomic DNAs, Pseudonocardia autotrophica strain was identified to contain a unique polyene-specific CYP gene. The genomic DNA library screening using the polyene-specific CYP gene probe revealed the positive cosmid clone containing an approximately 34.5 kb DNA fragment revealed a total of seven complete and two incomplete open reading frame (ORFs), which are highly homologous but unique to previously-known polyene biosynthetic genes. These results suggest that the polyene-specific screening approach should be an efficient way of isolating potectially-valuable cryptic polyene biosynthetic gene cluster from various rare actinomycetes.

• The Plant Pathology Journal
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• 제22권2호
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• pp.168-173
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• 2006

To develop the diagnostic method for the viral infection in apple, the partial genes corresponding to the N-terminal region of RNA polymerase of Apple stem grooving virus (ASGV) and coat protein of Apple chlorotic leaf spot virus (ACLSV) were characterized from the infected apple cultivars in Korea. Based on the nucleotide sequences of the characterized partial genes, the virus gene-specific primers were designed for the detection of ASGV and ACLSV infected in species of Malus. The RT-PCR using the primers for the genes of ASGV and ACLSV successfully gave rise to 404 and 566 bp DNA fragments, respectively. Using those viral gene-specific primers, the multiplex RT-PCR assays were also established to diagnose the mixed infection by ASGV and ACLSV simultaneously. Furthermore, the control primers, which have to be included for the RT-PCR as an internal control, were designed using the nucleotide sequence of the gene encoding elongation factor $1{\alpha}(EF1{\alpha})$. This multiplex RT-PCR including the control primers provides more reliable, rapid and sensitive assay for the detection of ASGV and ACLSV infected in Korean apple cultivars.