• 제목/요약/키워드: Specific primer

검색결과 875건 처리시간 0.028초

형태적.분자생물학적 방법에 의한 Phellinus linteus의 동정에 관한 연구 (Identification of Phellinus linteus by Morphological Characteristics and Molecular Analysis)

  • 김상희;김수호;성재모
    • 한국균학회지
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    • 제27권5호
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    • pp.337-340
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    • 1999
  • rDNA내의 ITS region의 염기서열 분석 결과를 토대로 19종의 Phellinus 속균중 Phellinus linteus를 특이적으로 동정할 수 있는 primer를 제작하였다. 이 특이 primer는 ITS1과 ITS2내에 위치하며 이들 spacer region에 인접해 있는 universal Primer내에 위치해 있다. 총 4개의 Primer(universal primer인 ITS-1F와 ITS-4 그리고 특이 primer인 PL-F와 PL-R)가 한국에서 채집된 Phellinus 속균중 Phellinus linteus를 동정하는데 사용되었다. Phellinus linteus의 증폭된 DNA크기는 800bp(ITS-1F/ITS-4)와 720 bp(ITS-1F/PL-R과 PL-F/ITS-4)에 해당하는 2개의 band, 그리고 610 bp(PL-F/PL-R)인 것으로 나타났다. 한국에서 채집된 23종의 Phellinus속균중 13종이 Phellinus linteus인 것으로 확인되었다.

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다래나무속 식물의 분류 및 계통 특이밴드 탐색을 위한 범용 프라이머 개발 (Development of Universal Primers for Phylogenetic Analysis and Species-specific Band Identification in the Genus Actinidia)

  • 김성철;장기창;송은영;김공호;정용환;김미선;오순자;고석찬
    • 한국자원식물학회지
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    • 제17권2호
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    • pp.107-115
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    • 2004
  • 참다래 육종을 위한 종 분류와 분자 표지인자로서 유용하게 이용될 수 있는 primer를 개발하기 위하여 참다래 genome 특이 반복 염기서열로부터 19∼20base 크기로 18개의 primer를 제작하여 kiwifruit target primer(KT primer)라 명명하였으며, 동아시아 지역에서 수집된 7종 22계통의 다래나무 속 식물을 이용하여 활용 가능성 을 조사하였다. 유연관계 분석을 위하여 7개의 primer가 선발되었으며, 이를 이용한 RAPD 결과 크게 2개의 군으로 나뉘어 졌다. 제 1 군(A. arguta, A. melanandra, A. kolumikta와 A. marcrosperma)은 주로 과실에 전혀 털이 없으며 잎에는 털이 전혀 없거나 어렸을 때 극소량의 연모가 있다가 없어지는 그룹으로서 Leiocarpae 절에 속하였다. 제 2 군(A. chinensis, A. deliciosa 및 A. eriantha)은 어린 과실에서는 털이 많았다가 성숙하면서 털이 없어지는 계통 및 잎과 줄기에 털이 아주 많거나 조밀한 솜털이 있는 그룹으로서 Stellatae절에 속하였다. 제 2 군은 Stellatae 절에서도 Pefectae 아절에 속하는 것으로 A. chinensis, A. deliciosa 및 A. eriantha가 포함되었으며, 다시 A. chinensis와 A. deliciosa를 포함하는 그룹과 A. eriantha 등 2개의 그룹으로 나뉘어졌다. 같은 부모로부터 유래된 것으로 알려진 A. chinensis와 A. deliciosa는 80%의 유사도에서 두 개의 그룹으로 나뉘어졌다. 또한 PCR 결과 A. deliciosa 종 및 헤이워드와 토무리 계통 특이 밴드가 KT12F와 KT6F에서 나타났으며, 유전양상 분석에서 KT7F와 KT12F가 유용하였다. 본 연구 결과 KT primer는 참다래의 유전양상 분석과 특이한 유전양상을 나타내는 개체선발 및 도태에 유용하게 이용될 수 있고, 또한 참다래 육종 효율향상에 많은 도움을 줄 수 있다고 판단되었다.

Selection of PCR Markers and Its Application for Distinguishing Dried Root of Three Species of Angelica

  • Jin, Dong-Chun;Sung, Jung-Sook;Bang, Kyong-Hwan;In, Dong-Su;Kim, Dong-Hwi;Park, Hee-Woon;Seong, Nak-Sul
    • 한국약용작물학회지
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    • 제13권2호
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    • pp.121-125
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    • 2005
  • An analysis of RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction) was performed with three Angelica species (A. gigas Nakai, A. sinensis (Olive.) Diels and A. acutiloba Kitag) in an effort to distinguish between members of these three species. Two arbitrary primers (OPC02, OPD11) out of80 primers tested, produced 17 species-specific fragments among the three species. Eight fragments were specific for A. sinensis, four fragments specific for A. gigas, five specific for A. acutiloba. When primers OPC02 and OPD11 were used in the polymerase chain reaction, RAPD-PCR fragments that were specific for each of the three species were generated simultaneously. Primer OPC02 produced eight species-specific fragments: four were specific for A. sinensis, one for A. gigas, and three for A. acutiloba. Primer OPD11 produced nine speciesspecific fragments: four for A. sinensis, three for A. gigas, and two for A. acutiloba. The RAPD-PCR markers that were generated with these two primers should rapidly identify members of the three Angelica species. The consistency of the identifications made with these species-specific RAPD-PCR markers was demonstrated by the observation that each respective marker was generated from three accessions of each species, all with different origins. We also performed the RAPD-PCR analysis with the dried Angelica root samples that randomly collected from marketed and from the OPC02 primer, obtained a A. gigasspecific band and the band were cloned and sequenced.

Genetic Similarity and Difference of Marsh Clam (Corbicula leana) Obtained by RAPD-PCR

  • Yoon, Jong-Man;Park, Hwan-ha;Choe, Sun-Nam
    • 한국어업기술학회:학술대회논문집
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    • 한국어업기술학회 2002년도 춘계 수산관련학회 공둥학술발표회
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    • pp.279-280
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    • 2002
  • Genomic DNA from the muscle of marsh clam (Corbicula leana) from Gochang was extracted in order to identify genetic differences and similarity by randomly amplified polymorphic DNAs-polymerase chain reaction. 3.28 of the 23.0 polymorphic bands per lane were found to be polymorphic in marsh clam. Also, about 4.34% of total polymorphic bands were either specific to marsh clam. The major common bands of 0.28 kb generated by primer OPB-15 (GGAGGGTGTT) were present in every individuals, respectively, which were polymorphic. This common bands which present in every individuals should be diagnostic of specific strains, species and/or their relatedness. Primer OPB-19 (ACCCCCGAAG) produced the highest number of specific bands, which was 12. The specific minor band of 0.07 kb was present in lane 22, which were polymorphic. Especially, only a specific band (1.35 kb) identifying individuals was observed in lane 22.

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Genetic Similarity and Difference of Marsh Clam (Corbicula leana) Obtained by RAPD-PCR

  • Yoon, Jong-Man;Park, Kwan-Ha;Choe, Sun-Nam
    • 한국양식학회:학술대회논문집
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    • 한국양식학회 2002년도 춘계 한국양식학회 학술대회 발표요지
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    • pp.171-172
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    • 2002
  • Genomic DNA from the muscle of marsh clam (Corbicula leana)from Gochang was extrected in order to identify genetic differences and similarity by randomly amplified polymorphic DNAs-polymerase chain reaction. 3.28 of the 23.0 polymorphic bands per lane were found to be polymorphic in marsh clam. Also, about 4.34% of total polymorphic bands were either specific to marsh clam. The major common bands of 0.28 kb generated by primer OPB-15 (GGAGGGTGTT) were present in every individuals, respectively, which were polymorphic. This common bands which present in every individuals should be diagnostic of specific strains, species and-or their relatedness. Primer OPB-19 (ACCCCCGAAG) produced the highest number of specific bands, which was 12. The specific minor band of 0.07 kb was present in lane 22, which were polymorphic. Especially, only a specific band (1.35kg) identifying individuals was observed in lane 22.

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Development of Penicillium italicum-Specific Primers for Rapid Detection among Fungal Isolates in Citrus

  • Chen, Kai;Tian, Zhonghuan;Jiang, Fatang;Long, Chao-an
    • Journal of Microbiology and Biotechnology
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    • 제29권6호
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    • pp.984-988
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    • 2019
  • Blue mold in citrus is caused by Penicillium italicum. In this study, the P. italicum-specific primers were developed for rapid detection based on the conserved genes RPB1 and RPB2 among Penicillium genomes. The two primer pairs RPB1-a and RPB1-b proved to be specific to detect P. italicum. The PCR assay among 39 fungal isolates and the colonial, pathogenic morphologies and molecular methods validated the specificity and reliability of these two primer pairs. This report provided a method and P. italicum-specific primers, which might greatly contribute to citrus postharvest industry.

Genetic Distances of Three Mollusk Species Investigated by PCR Analysis

  • Oh, Hyun;Yoon, Jong-Man
    • 한국발생생물학회지:발생과생식
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    • 제18권1호
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    • pp.43-49
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    • 2014
  • Three species of Nortamea concinua (NC) and Haliotis discus hannai (HDH) from Tongyeong and Sulculus diversicolor supertexta (SDS) are widely distributed on the coast of the Yellow Sea, southern sea and Jeju Island in the Korean Peninsula under the innate ecosystem. There is a need to understand the genetic traits and composition of three mollusk species in order to evaluate exactly the patent genetic effect. PCR analysis was performed on DNA samples extracted from a total of 21 individuals using seven decamer oligonucleotides primers. Seven primers were shown to generate the unique shared loci to each species and shared loci by the three species which could be clearly scored. A hierarchical clustering tree was constructed using similarity matrices to generate a dendrogram, which was facilitated by the Systat version 10. 236 specific loci, with an average of 56.3 per primer, were identified in the NC species. 142 specific loci, with an average of 44.7 per primer, were identified in the HDH species. Especially, 126 numbers of shared loci by the three species, with an average of 18 per primer, were observed among the three species. Especially, the decamer primer BION-75 generated 7 unique loci to each species, which were identifying each species, in 700 bp NC species. Interestingly, the primer BION-50detected 42 shared loci by the three species, major and/or minor fragments of sizes 100 bp and 150 bp, respectively, which were identical in all samples. As regards average bandsharing value (BS) results, individuals from HDH species (0.772) exhibited higher bandsharing values than did individuals from NC species (0.655). In this study, the dendrogram obtained by the seven decamer primers indicates three genetic clusters: cluster 1 (CONCINNA 01~CONCINNA 07), cluster 2 (HANNAI 08~HANNAI 14), cluster 3 (SUPERTEXTA 15~SUPERTEXTA 21). Comparatively, individuals of HDH species were fairly closely related to that of SDS species, as shown in the hierarchical dendrogram of genetic distances.

16S rDNA sequence에 대한 종특이성 primer를 이용한 중합효소연쇄반응증폭에 의한 Porphyromonas endodontalis의 동정에 관한 연구 (A STUDY ON THE IDENTIFICATION OF Porphyromonas endodontalis BY PCR USING SPECIES SPECIFIC PRIMERS FOR THE 16S rDNA)

  • 엄승희;임성삼;배광식
    • Restorative Dentistry and Endodontics
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    • 제24권1호
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    • pp.13-25
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    • 1999
  • P. endodontalis which was known to be associated with the infected root canals and periapical lesions is very difficult to detect by culture methods or traditional methods. Detection of bacteria using polymerase chain reaction(PCR) for 16S ribosomal DNA(rDNA) is fast, simple, and accurate with relatively small amount of target cells. 16S rDNA consist of conserved regions those are same to all species, and variable regions which represent species specificity. The 16S rDNA sequences of P. endodontalis and P. gingivalis were aligned and two highly variable regions were selected as a pair of species specific oligonucleotide primers for P. endodontalis. And then the pair of primers for PCR amplification was synthesized to identify P. endodontalis. The sequences of the species specific primers for the 16S rDNA of P. endodontalis were as follows ; sense primer[endo1]: 5'-CTATATTCTTCTTTCTCCGCATGGAGGAGG-3' antisense primer[endo2]: 5'-GCATACCTTCGGTCTCCTCTAGCATAT-3' In this study, for the identification of P. endodontalis without culture from the mixed clinical samples, PCR was done with species specific primers for the 16S rDNA sequences of P. endodontalis. The results were as follows : 1. The species specificity of the primers for the 16S rDNA of P. endodntalis was determined by the PCR methods. About 490bp amplicon which was specific only for P. endodntalis was produced with P. endodontalis. No amplicon was produced by PCR with other strains similar to P. endodontalis. 2. The synthesized species specific primers reacted with conventionally identified P. endodontalis which we have in conservative dentistry laboratory. 3. The identification of P. endodontalis using PCR technique with samples collected from infected root canals or periapical lesions was more sensitive than that of culture methods. 4. Seven samples revealed including P. endodontalis by PCR technique. Five of them were related with pains, two of them with sinus tract, three of them with foul odor, and three of them with purulent drainage. P. endodontalis was shown to have great relation with pains.

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PepN과 16S rRNA Gene Sequence 및 PCR 방법을 이용한 김치 젖산균의 동정 (Genetic Identification of the Kimchi Strain Using PCR-based PepN and 16S rRNA Gene Sequence)

  • 이명기;박완수;이병훈
    • 한국식품과학회지
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    • 제32권6호
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    • pp.1331-1335
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    • 2000
  • 김치 젖산균인 WL6는 API kit 또는 Biolog system방법에 의하여 동정한 결과, Leuconostoc mesenteroides ssp. cremoris, Leu. mesenteroides ssp. dextranicum 또는 Lactobacillus bifermentans로 나타나 동정되지 않았다. 그러나, pepN gene과 16S rRNA gene으로부터 2개의 specific-sequence primer set을 제조하여 PCR 방법으로 증폭한 후에 표준균주들과 비교한 결과, WL6는 Lactobacillus bifermentans로 추정되었다.

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Quantitative Analysis of Human- and Cow-Specific 16S rRNA Gene Markers for Assessment of Fecal Pollution in River Waters by Real-Time PCR

  • Jeong, Ju-Yong;Park, Hee-Deung;Lee, Kyong-Hee;Hwang, Jae-Hong;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • 제20권2호
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    • pp.245-253
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    • 2010
  • The base sequences representing human- and cow-specific 168 rRNA gene markers identified in a T-RFLP analysis were recovered from clone libraries. The human- and cow-specific primers were designed from these sequences and their specificities were analyzed with fecal DNAs from human, cow, and pig. The AllBac primer set showed positive results for all human, cow, and pig samples, whereas the human-specific primer set showed positive result only for the human sample but not for the cow or pig samples. Likewise, the cow-specific primer set showed positive results only for the cow sample but not for the human or pig samples. Real-time PCR assay with these primers was developed for the identification and quantification of fecal pollution in the river water. The human- and cow-specific markers were detected in the order of 9 $\log_{10}$ copies per gram wet feces, which were two orders of magnitude lower than those of total Bacteroidales. For the river water samples, the human-specific marker was detected in $1.7-6.2\;\log_{10}$ copies/100 ml water, which was 2.4-4.9 orders of magnitude lower than those of total Bacteroidales. There was no significant correlation between total Bacteroidales and conventional fecal indicators, but there was a high correlation between Bacteroidales and the human-specific marker. This assay could reliably identify and quantify the fecal pollution sources, enabling effective measures in the watersheds and facilitating water quality management.