• Title/Summary/Keyword: RFLP.

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Genotypic Characterization of Cherry Witches' Broom Pathogen Taphrina wiesneri Strains (벚나무 빗자루병균 Taphrina wiesneri의 유전적 특성)

  • Seo, Sang-Tae;Jeong, Su-Jee;Lee, Seung-Kyu;Kim, Kyung-Hee
    • Research in Plant Disease
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    • v.17 no.1
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    • pp.99-101
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    • 2011
  • The ascomycetous fungus Taphrina wiesneri, the pathogen of cherry witches' broom, is highly pathogenic to Prunus yedoensis, the most widely planted cherry trees in Korea as park and roadside trees. A collection of 13 strains of the pathogen in Korea and Japan was characterized by 18S rDNA gene sequence and restriction fragment length polymorphism (RFLP) analysis. In cluster analysis based on 18S rDNA gene sequence the strains were divided into 2 clusters. In RFLP analysis of the rDNA-IGS region using HhaI, the strains were separated into four patterns, B, C, D and G, of which pattern G was new.

Comparison of Macroscopic Inspection and Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) for the Detection of Anisakis simplex complex (고래회충 검출을 위한 육안검사법과 중합효소연쇄반응-제한효소절편길이다형성의 비교)

  • Kang, Ju-Hee;Lee, Min-Hwa;Lee, Kang-Bum;Choi, Chang-Sun
    • Journal of Food Hygiene and Safety
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    • v.23 no.4
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    • pp.314-318
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    • 2008
  • This research aimed to compare the detection methods of Anisakis simplex in Sea fish by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and macroscopic inspection. We examined 18 Trichiurus lepturus, 11 Scomber japonicus, and 65 Todarodes pacificus collected from the retail markets in the areas of Uljin, Kyuonggi province and Seoul. As the result of examinations, we found that detection rate of Anisakis simplex by macroscopic observation was 89% in Trichiurus lepturus, 90.9% in Scomber japonicus, 32.3% in Todarodes pacificus. The detection rate of Anisakis simplex by PCR-RFLP was 77.7% in Trichiurus lepturus, 81.8% in Scomber japonicus, 26.1% in Todarodes pacificus. We could conclude that PCR-RFLP method of Anisakis simplex was more specific rather than macroscopic observation.

Cytochrome B 유전자의 PCR-RFLP Marker에 의한 식육자원의 축종 판별

  • Sin, Seong-Cheol;Choe, Eun-Ju;Heo, Yeon-Beom;Baek, Myeong-Gi;Gwon, Su-Yeon;Kim, Bo-Hyeon;Jeong, Gu-Yong;Jeong, Ui-Ryong
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2005.05a
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    • pp.195-198
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    • 2005
  • 본 연구는 cytochrome B 유전자의 PCR-RFLP 분석기법을 이용하여 다양한 식육자원 및 각종 가공 육제품의 원료육에 대한 정확하고 재현성 높은 축종 및 육종 감별기술을 개발하기 위하여 수행되었다. 국내에서 유통되고 있는 7종류 축종(닭, 양, 돼지, 소, 사슴, 말, 염소)의 육류로부터 cytochrome B 유전자의 특정 염기서열을 포함하는 primer를 설계 제작하여 PCR-RFLP 분석을 실시하였다. 각 축종의 근육조직으로부터 genomic DNA를 추출하고 PCR 증폭 반응을 수행한 후 얻어진 PCR 증폭산물(359 bp)을 두 종류의 제한효소(Hae Ш 및 Hinf I)로 각각 절단한 결과 축종 간 그리고 제한효소 간에 명확한 차이를 보이는 종 특이적인 PCR-RFLP marker를 검출하였다. 따라서, 본 연구에서 개발한 cytochrome B 유전자의 종 특이적 PCR-RFLP marker는 각종 식육 및 가공 육제품의 육종 및 축종 판별에 매우 유용한 DNA marker로 이용될 수 있을 것이다.

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PCR-RFLP 분석기법을 이용한 가금육의 종(닭, 칠면조, 오리) 판별 기술 개발

  • Sin, Seong-Cheol;Choe, Eun-Ju;Lee, Jun-Je;Kim, Hui-Seon;Jo, Ha-Na;Jeon, Sang-Hui;Jeong, Gu-Yong;Jeong, Ui-Ryong
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2005.05a
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    • pp.199-202
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    • 2005
  • 본 연구는 cytochrome B 유전자의 PCR-RFLP 분석기법을 이용하여 다양한 가금류 식육자원 및 각종 가공 육제품의 원료육에 대한 정확하고 재현성 높은 축종 및 육종 감별기술을 개발하기 위하여 수행되었다. 국내에서 유통되고 있는 종간 근연관계가 높은 3종의 가금육(닭, 칠면조 및 오리)의 육류에서 분리한 DNA 시료를 대상으로 cytochrome B 유전자의 특정 염기서열을 포함하는 primer를 설계 제작하여 PCR-RFLP 분석을 실시하였다. 각 축종의 근육조직으로부터 genomic DNA를 추출하고 PCR 증폭 반응을 수행한 후 얻어진 PCR 증폭산물(359 bp)을 두 종류의 제한효소(HaeШ 및 Hinf I)로 각각 절단한 결과 특히, HaeШ 제한효소에서 가금류의 축종 간 그리고 제한효소 간에 명확한 차이를 보이는 종 특이적인 PCR-RFLP marker를 검출하였다. 따라서, 본 연구에서 개발한 cytochrome B 유전자의 가금류 종 특이적 PCR-RFLP marker는 가금류의 식육 및 가공 육제품의 육종 및 축종 판별에 매우 유용한 DNA marker로 이용될 수 있을 것이다.

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Characterization of infectious bursal disease viruses isolated in Korea using RT/PCR and RFLP analysis (RT/PCR과 RFLP 분석에 의한 Infectious bursal disease virus(국내분리주)의 특성 규명)

  • Kwon, Hyuk-moo;Kim, Dae-kyu;Seong, Hwan-woo
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.104-110
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    • 1999
  • Field infectious bursal disease viruses (IBDVs) were isolated from IBDV-suspected commercial chickens. The variable region in VP2 gene of six Korean IBDV isolates (K-IBDVs) and IBD vaccines was examined using the reverse transcriptase / polymerase chain reaction-restriction fragment length polymorphism (RT/PCR-RFLP) assay. With all K-IBDVs and vaccine IBDVs, a 474-bp fragment of the VP2 gene was amplified and tested with various restriction enzymes. Restriction enzymes BstNI and StyI differentiated K-IBDV isolates and IBD vaccines into four groups. Restriction enzyme profiles of K-IBDV isolates were different from them of IBD vaccines. K-IBDV isolates except for 310 isolate had specific SspI and TaqI recognition sites, which were recognized in highly virulent IBDVs, but IBD vaccines had no those sites. This study showed that RT/PCR-RFLP assay was thought to be valuable tool for differentiation of IBDVs and identification of highly virulent IBDV.

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Analyses of Genetic Relationships of Collectorichum spp. Isolated from Sweet Persimon with RAPD and PCR-RFLP. (단감나무로부터 분리한 탄저병 병원균 Colletotrichum spp.의 RAPD와 PCR-RFLP를 이용한 유연관계 분석)

  • 김희종;엄승희;이윤수
    • Korean Journal of Microbiology
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    • v.38 no.1
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    • pp.19-25
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    • 2002
  • Colletotrichum species are important fungal pathogen that cause great damages on various host plant species worldwide. In Korea, Colletotrichum species cause massive economic losses on apple, peach, grape, and essecially, sweet persimon productions. In the past, Identification of the pathogen and the studies on the genetic relationships among the pathogenic isolates were mainly based on morphology, cultural characteristics, and the difference in pathogenicity. However, in recent years, these traditional methods have been replaced with molecular methods to solve the difficulty of classification on pathogens. Therefore, in this study, RAPD and PCR-RFLP methods were employed for the studies of genetic relationship among the different isolates of Colletotrichum species that cause damages on sweet persimon. As a results of genetic relationship analysis, Colletotrichum species tested were divided into two big groups or five small groups.

Discrimination of Hanwoo from Holstein/black Angus meat by PCR-RFLP of MC1R gene (MC1R 유전자의 PCR-RFLP를 이용한 한우육과 젖소육/black Angus 수입육의 구분)

  • Kim, Tae-Jung;Lee, Jae-Il
    • Korean Journal of Veterinary Research
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    • v.45 no.3
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    • pp.335-339
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    • 2005
  • The melanocortin 1 receptor (MC1R) plays an important role in regulation of melanin pigment synthesis within mammalian melanocytes. Mutations within the gene encoding MC1R have been shown to explain coat color variations within several mammalian species including cattle. To develope a rapid and accurate method for the identification of Hanwoo, we performed a modified PCR-RFLP analysis of MC1R gene using single nucleotide polymorphism (SNP) within MC1R as a target. A size of 538 bp (537 bp for Hanwoo) was amplified by PCR, digested with Hpa II, and electrophoresed on a 1.5% agarose gel. A PCR product from Hanwoo showed a single band of 537 bp, whereas two fragments of 328 bp and 210 bp were detected in both Holstein and Black angus. The current result suggests that the PCR-RFLP using our primers and enzyme digestion system would be very accurate, easy and reproducible method to discriminate between Hanwoo and Holstein/Black angus meat.

Identification of Meat Species Using PCR-RFLP Marker of Cytochrome b Gene

  • Shin, Sung-Chul;Chung, Ku-Young;Chung, Eui-Ryong
    • Food Science of Animal Resources
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    • v.26 no.3
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    • pp.375-379
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    • 2006
  • Food labeling regulations require that the meat species in various meat products are accurately declared to the consumer. Substitution or adulteration of costly meat with a cheaper one is one of the most common problems in the meat industry. In this study, PCR-restriction fragment length polymorphism(RFLP) method of the mitochondrial cytochrome b(mt cyt b) gene has been applied for identification of the origin of six mammalian meat species(beef, port horse, goat, mutton and deer) and three poultry meat species(chicken, turkey and duck) as raw materials for meat products. PCR was used to amplify a variable region of mt cyt b gene. Meat species differentiation was determined by digestion of the amplified products with a 359 bp fragment using HaeIII and HinfI restriction enzymes, which generated species-specific RFLP patterns. This PCR-RFLP DNA marker of mt cyt b gene could be very useful for the accurate and reliable identification and discrimination of animal meat species in routine analysis.

RFLP Analysis of the mtDNA COI Region in Four Abalone Species

  • Park, Choul-Ji;Kijima, Akihiro
    • Fisheries and Aquatic Sciences
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    • v.9 no.3
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    • pp.101-106
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    • 2006
  • The cytochrome c oxidase subunit I (COI) gene region of mitochondrial DNA (mtDNA) was examined in four abalone species to estimate its utility as a genetic marker using restriction fragment length polymorphism (RFLP) analysis. The utility was evaluated in terms of genetic divergence and relationships among Haliotis discus hannai, H. rufescens, H. rubra, and H. midae in both hemispheres of the world. There was clear genetic divergence in the mtDNA COI region between all pairs of the four species. Moreover, relationships among the abalone species were reflected in their geographical distributions and morphological characteristics. Therefore, RFLP analysis of the mtDNA COI region is a suitable genetic marker for the estimation of genetic divergence and relationships among abalone species. However, it is not effective for the evaluation of genetic differences within abalone species.