• 제목/요약/키워드: polymorphic bands

검색결과 282건 처리시간 0.029초

Susceptibility Testing of Helicobacter pylori to Metronidazole and DNA Fingerprints of Resistant Strains in Singapore

  • HUA, JIESONG;KHAY-GUAN YEOH;PENGYUAN ZHENG;HAN CHONG NG;BOW HO
    • Journal of Microbiology and Biotechnology
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    • 제9권3호
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    • pp.328-333
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    • 1999
  • Susceptibility of 61 strains of Helicobacter pylori to metronidazole was examined by both the disk diffusion method using a cut-off of 15㎜ for resistance and the E test with a cut-off of 8㎎/l. The MIC/sub 50/ and MIC/sub 90/ by the E test were 2 ㎎/l and 256㎎/l, respectively. Metronidazole resistance was found in 22 (36%) out of the 61 H. pylori strains by the E test and in three additional strains by the disk diffusion method. Amongst the latter three isolates, the MICs by the E test were 4 ㎎/l, 6㎎/l, and 6㎎/l, respectively. These figures are one log₂ or half log₂ dilution lower than the cut-off of 8㎎/l recommended as resistance for the E test. All 22 metronidazole resistant H. pylori isolates by the E test that were subjected to random amplified polymorphic DNA (RAPD) fingerprinting showed different DNA fingerprints. Interestingly, >90% of resistant isolates possess two common DNA bands of 0.4 and 0.9 kb. This study demonstrates that the results of the disk diffusion method for testing H. pylori susceptibility to metronidazole correlates well with that of the E test. The criteria for interpretation need to be internationally standardized so that the results from different centers can be compared.

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해조류로부터 Arbitrary 및 ITS Primer들을 사용한 직접 PCR 유전자 증폭반응의 한계 (Limits of Direct PCR Amplification from Seaweeds Using Arbitrary and ITS Primers)

  • 김용국;진형주;박선미;진덕희;홍용기
    • 생명과학회지
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    • 제9권1호
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    • pp.15-21
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    • 1999
  • PCR 기법을 응용한 RAPD 방법은 대상 생물의 유전정보를 전혀 모르는 상태에도 arbitrary primer를 이용하여 증폭함으로써 생물 종간의 유전적 표식자를 확인할 수 있는 간편하고 유익한 유전분석 방법이다. 이같은 RAPD 방법을 이용하여 해조류 방사무늬 김, 미역, 구멍갈파래에 대하여 DNA를 추출하지 않고 직접 생 엽체 및 생 사상체, 생 배우체를 PCR template로 사용하여 PCR product를 생산하였다. 그러나 specific primer를 이용한 nulear r DNA의 internal trancribed spacer (ITS) 부위는 생성물을 만들지 못하였다. 간편한 LiCl 방법에 의하여 추출된 DNA를 사용하였을 때는 IST 및 RAPD 모두 PCR product를 생산하였다. 방사무늬 김의 엽체 (haploid)와 사상체 (dip-loid)로 부터의 ITS 생성물은 동일하였으나 RAPD 생성물은 사용한 arbitrary primer에 따라 36-50$\%$의 다른 band가 만들어졌다. 또한 직접 생 조직을 사용하였을때와 추출 DNA를 사용하였을 때도 53-57$\%$의 다른 band가 만들어 줬다. 그러므로 RAPD 방법으로 유전분석 실험에는 동일한 ploidy의 조직을 template로서 사용하는 것이 중요하다.

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Evaluation of Genetic Diversity among Soybean Genotypes Using SSR and SNP

  • Lee, Suk-Ha;P. Tanya;O, Srinives;T. Toojinda;A. Vanavichit;Ha, Bo-Keun;Bae, Jeong-Suk;Moon, Jung-Kyung
    • 한국작물학회지
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    • 제46권4호
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    • pp.334-340
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    • 2001
  • Two different types of molecular markers, simple sequence repeat (SSR) and single nucleotide polymorphism (SNP), were used to measure genetic diversity among five Korean, eight Thai, and three wild soybeans. For SSR analysis, a total of 20 markers were surveyed to detect polymorphisms. For SNP analysis, four primers were designed from consensus sequence regions on disease resistance protein homolog genes, and used to amplify the genomic region. The PCR products were sequenced. A number of polymorphic SSR and SNP bands were scored on all genotypes and their genetic similarity was measured. Clustering analysis was performed independently on both types of markers. Clustering based on SSR markers separated the genotypes into three main groups originated from Korea, Thailand, and wild soybeans. On the other hand, two main groups were classified using SNP analysis. It seemed that SSR was more informative than SNP in this study. This may be due to the fact that SNP was surveyed on the smaller genomic region than SSR. Grouping based on the combined data of both markers revealed similar results to that of SNP rather than that of SSR. This might be due to the fact that more loci from SNP were considered to measure genetic relatedness than those from the SSR.

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동위효소와 RAPD 법을 이용한 제주 자생 새우란, 금새우란, 왕새우란의 근연관계 분석 (Isozyme and Randomly Amplified Polymorphic DNA (RAPD) Analysis for Genetic Relationship among Calanthe discolor, C. sieboldii, and C. bicolor Native to Cheju Island)

  • 현명력;최지용;서정남;소인섭;이종석
    • 원예과학기술지
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    • 제17권2호
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    • pp.141-143
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    • 1999
  • 제주 자생 새우란, 금새우란, 왕새우란의 분류를 위한 동위효소 분석과 RAPD법을 이용한 유전적 근연관계 조사 결과, peroxidase와 esterase의 동위효소(isozyme)분석에서 왕새우란의 밴드양상은 새우란과 금새우란이 동시에 나타나는 밴드에서만 밴드가 나타났다. RAPD를 이용한 유전적인 근연관계에서 왕새우란은 새우란과 금새우란 모두와 비교적 가까웠지만 새우란과 금새우란의 상호간 근연관계는 가장 멀었다. 즉, 왕새우란은 유전적인 근연관계에서 새우란과 금새우란의 중간적인 위치를 나타내었다. 따라서, 왕새우란은 새우란과 금새우란의 자연적인 교잡으로 출현된 새로운 종으로 사료된다.

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Analysis of Genetic Diversity of Korean Accessions of the Genus Acorus Using RAPD Markers and NIR Spectroscopy

  • Lee, Ja-Hyun;Kim, In-Seon;Lee, Seong-Gene;Rim, Kwang-Sub;Kim, Sung-Gil;Han, Tae-Ho
    • 원예과학기술지
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    • 제29권3호
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    • pp.232-239
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    • 2011
  • The genus Acorus is known as an indigenous medicinal plant. Genetic diversity of thirteen accessions of A. calamus and eight of A. gramineus, with an accession of Colocasia antiquorum and two of Iris pseudacorus as outgroups, were evaluated using RAPD markers for cluster analysis and principal coordinate analysis, and NIR spectroscopic profiles for principal component analysis.A total of 371 polymorphic bands were obtained by using the selected 12 random primers. The genetic distances were estimated from 0.03 to 0.31 within A. calamus and from 0.03 to 0.51 within A. gramineus. The dendrogram and three-dimensional plot separated the accessions into four distinct groups (A. calamus, A. gramineus, C. antiquorum, and I. pseudacorus). Moreover, for the diversity among genus Acorus, eleven A. calamus accessions, one A. gramineus accession, and two I. pseudacorus accessions were non-destructively analyzed from their leaves by NIR spectroscopy, which discriminated Acorus accessions like the RAPD analysis. Interestingly, thirteen accessions of A. calamus were clustered into two groups based on RAPD and NIR analyses, which indicates that there are two ecotypes of A. calamus in Korea. An accession (CZ) of A. calamus with yellow stripe on leaves was closely grouped with another (CX) at a genetic distance (GD) of 0.03, which shows that the stripe trait might be generated by chimeric mutation. The genetic distance between A. calamus and A. gramineus was revealed to be farthest from 0.80 to 0.88 GD. In genus Acorus the genetic diversity and genetic variation were identified by using RAPD marker technique and non-destructive NIRs.

RAPD 분석에 의한 여수 돌산갓의 유연관계 분석 (Genetic Relationship Based on RAPD Analysis of Yeosu Dolsan Leaf Mustard(Brassica juncea))

  • 이인호;박종인;정운섭;정효진;;노일섭
    • 생명과학회지
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    • 제20권1호
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    • pp.66-70
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    • 2010
  • RAPD 방법을 이용하여 14의 돌산갓 품종의 유전적 다형성을 분석하였다. 39개의 random primer를 실험하여 그 중 7개의 다형성을 나타내는 primer를 선발 하였다. 증폭된 PCR 산물은 0.2~3.5 kb 사이에서 재현성 있는 밴드를 나타내었으며, 각 primer에 의해 증폭된 밴드의 수는 2~27개로 다양하였고, 평균 8.7개였다. UPGMA 분석에 의해 14개 품종은 3개의 그룹으로 나뉘었다. 이 결과들로부터 갓의 유전적 다양성 검토 및 $F_1$ 품종 생산을 위한 교배 조합 작성에 RAPD 분석은 유용하다는 것을 나타내었다.

Analysis of Genetic Relationship Among Native Pears Grown in Korea and Several Commercially Developed Cultivars from Two Pyrus Species Based on RAPD Analysis

  • Cho, Dong-Wook;Oh, Jin-Pyo;Chung, Kyu-Hwan
    • 한국자원식물학회지
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    • 제20권6호
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    • pp.563-569
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    • 2007
  • RAPD analysis showed that all the OTUs of 'Sandolbae' were the same species because amplified band patterns of all samples generated by each of 5 random primers were identical. Even though there were different environmental conditions, all the "Chuiangne" trees from three different places were the same species, and also all the "Cheongshilli" trees were the same species too. No genetic variations were detected between native Korean pears grown in the habitats and in the research field. Because 212 polymorphic bands were generated by 9 primers selected through primer screening, they were possible to analyze genetic relationship among naturally growing three native Korean pears and nine cultivars of Pyrus pyrifolia and P. communis. Based on the RAPD analysis, three main groups were formed. The first group represented the Six P. pyrifoia cultivars, the second group was the three native Korean pears, and the last group was the three P. communis cultivars. Genetic distance between 'Wonwhang' and 'Chojuro' was closer than other cultivars in group 1 since dissimilarity index value between these two cultivars was 50.82. However, genetic distance between 'Niitaka' and 'Chojuro' was the most distant compared to the others in group 1. In group 2, 'Sandlobae' was genetically closer to 'Chuiangne' than 'Cheongshilli' because dissimilarity index value between 'Sandlobae' and 'Chuiangne' was smaller, 50.82, than the value between 'Sandlobae' and 'Cheongshilli', 63.636. In group 3, 'Old Home' was genetically closer to 'Bartlett' than 'Kaiser Alexander(or Bosc)'. Group 3 composed of P. communis cultivars was genetically further than other two groups, P. pyrifolia cultivars and native Korean pears.

Vegetative Growth and Phylogenetic Relationship of Commercially Cultivated Strains of Pleurotus eryngii based on ITS sequence and RAPD

  • Alam, Nuhu;Shim, Mi-Ja;Lee, Min-Woong;Shin, Pyung-Gyun;Yoo, Young-Bok;Lee, Tae-Soo
    • Mycobiology
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    • 제37권4호
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    • pp.258-266
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    • 2009
  • Pleurotus eryngii, known as king oyster mushroom has been widely used for nutritional and medicinal purposes. This study was initiated to screen the suitable conditions for mycelial growth and to determine the phylogenetic relationship of the selected strains. Optimal mycelial growth was observed at $30{^{\circ}C}$ and minimum mycelial growth observed at $10{^{\circ}C}$. This mushroom tolerates a broad pH range for mycelial growth, with most favorable growth observed at pH 6. Results also indicated that glucose peptone, yeast malt extract and mushroom complete media were favorable growth media, while Hennerberg and Hoppkins media were unfavorable. Dextrin was the best and xylose the least effective carbon sources. Results revealed that inorganic nitrogen sources were less effective than organic sources for the mycelial growth of P. eryngii. Investigation of genetic diversity is necessary to identify the strains. The ITS region of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 214 to 222 bp and 145 to 236 bp, respectively. The sequence of ITS2 was more variable than that of ITS1, and the 5.8S sequences were identical. A phylogenetic tree based on the ITS region sequences indicated that selected strains could be classified into six clusters. Fourteen IUM and ATCC- 90212 strains were also analyzed by RAPD with 20 arbitrary primers. Fourteen of these primers were efficiently amplified the genomic DNA. The number of amplified bands varied with the primers and strains, with polymorphic fragments in the range from 0.2 to 2.3 kb.

Molecular Identification and Effects of Temperature on Survival and Growth of Hybrids between Haliotis gigantea Gmelin (♀) and Haliotis discus hannai Reeve (♂)

  • An, Hye Suck;Han, Jong Won;Hwang, Hyun-Ju;Jeon, Hancheol;Jung, Seung-Hyun;Jo, Seonmi;Choi, Tae-Young;Hyun, Young Se;Song, Ha Yeun;Whang, Ilson
    • 한국해양생명과학회지
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    • 제2권2호
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    • pp.83-89
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    • 2017
  • In abalones, interspecific hybridization has been suggested as a possible means to increase production and desired traits for the industry. In Korea, Haliotis gigantea is considered a species with a larger size and higher temperature tolerance than H. discus hannai. However, H. discus hannai is considered the most valuable and popular fishery resource due to its better acceptance and higher market prices. Thus, viable interspecific hybrids have been produced by artificial inseminating H. gigantea eggs with H. discus hannai sperm. However, the reciprocal hybrid cross was not successful. In this study, the hybridity and the growth and thermal tolerance performance of the interspecific hybrids were examined. A combination of various assays revealed maximum growth occurrence at 21℃ and the higher growth rate in the hybrids than that of H. discus hannai parent. In addition, the growth and survival at high-temperature (28℃) of the hybrids was equivalent to that of the highly tolerant H. gigantea parent, suggesting new possibilities to overcome the mass mortality in H. discus hannai during high temperature periods of summer season in Korea. Furthermore, the induced interspecific hybrid status was confirmed by the presence of species-specific bands for each parental species of the random amplified polymorphic DNA (RAPD) profiles using universal rice primer (URP), which could be used as speciesspecific markers to distinguish the hybrids and their parental species.

한국재래돼지의 G-, C-, 및 NOR-banding (G-, C-, and NOR-banding of Korean Native Pig Chromosomes)

  • 손시환;권오섭;백규흠;정원;조은정;강민영
    • Journal of Animal Science and Technology
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    • 제45권6호
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    • pp.901-910
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    • 2003
  • 본 연구는 한국재래돼지 염색체의 핵형 제시를 위하여 G-banding, C-banding 및 AgNORs를 분석하였다. 시험에 공시된 공시축은 축산기술연구소에서 선발 육종중인 재래돼지 종모돈 50두를 대상으로 각 개체별 혈액채취로서 혈액배양을 이용한 핵형 분석을 수행하였다. 한국재래돼지의 핵형은 38, XX 또는 XY로서 5쌍의 submetacentric chromosomes(Group I), 짧은 단완을 가진 2쌍의 acrocentric chromosomes(Group II), 5쌍의 metacentric chromosomes(Group III) 및 동원체가 말단부에 있는 6쌍의 acrocentric chromosomes(Group IV)로 구성된 36개의 상 염색체와 metacentric인 XX 또는 XY 성 염색체로 구성되어 있다. 재래돼지의 G-banding은 각 상동 염색체별 고유한 특징적 밴드 양상을 나타내고 있으며, 전체 염색체의 형태적 특징이나 대표적 landmarks는 국제표준핵형과 큰 차이가 없는 양상이다. 그러나 재래돼지의 경우 국제표준핵형에 비하여 보다 많은 band가 출현하였고 특히 1번, 3번, 5번, 6번, 7번, 8번, 13번, 14번, 15번, 16번, 17번, 18번 및 X 염색체에서 sub-bands의 분리를 나타내었다. 재래돼지의 C-banding은 비록 각 염색체들 간 heterochromatin의 양적 다형성이 존재하지만 거의 모든 상염색체의 동원체 부위에 C-bands가 나타나고, Y 염색체는 염색체의 전장에 걸친 heterochromatin의 분포를 보였다. AgNOR 염색에 의한 재래돼지의 NORs는 8번 및 10번 염색체의 동원체 부위에 확인되었고, 세포 당 NORs의 수는 2개에서 4개까지 관찰되었으며, 평균 2.13개로 분석되었다. 10번 염색체의 경우 모든 상동염색체에서 NORs가 나타나나 8번 염색체에서는 수적 다형성뿐만 아니라 양적 다형성을 나타내었다. 품종 간 NORs의 비교 분석에서 재래돼지의 NORs 수가 Yorkshire에 비해 유의적으로 높게 나타나 돼지의 NORs 분포 양상은 특히 8번 염색체에 있어 품종 간, 개체 간 및 세포 간에 다형적 변이 양상이 존재하는 것으로 사료된다.