• Title, Summary, Keyword: RFLP

Search Result 969, Processing Time 0.039 seconds

Differentiation of Vibrio spp. including Core Group Species by PCR-RFLP (PCR-RFLP에 의한 Vibrio core group을 포함한 Vibrio 종의 구분)

  • Park, Jin-Sook
    • Journal of Life Science
    • /
    • v.22 no.2
    • /
    • pp.245-250
    • /
    • 2012
  • The 16S rDNA - RFLP types for six Vibrio species (V. fluvialis, V. proteolyticus, V. vulnificus, V. mimicus) including two core group members, V. alginolyticus and V. parahaemolyticu s, and Grimontia (Vibrio) hollisae were determined using PCR-RFLP analysis. Six tetrameric restriction enzymes (Alu I, Cfo I, Dde I, Hae III, Msp I, and Rsa I) were selected for RFLP analysis. V. alginolyticus, V. parahaemolyticus, and V. proteolyticus showed the same RFLP pattern following digestion with four of the six used restriction enzymes: CfoI, DdeI, MspI, and RsaI. Various restriction enzyme combinations generated digests recognizable as distinct RFLP types for each of the assayed Vibrio species. In particular, AluI single digestion produced species specific band patterns that enabled the differentiation between these Vibrio species. Dendrogram based on restriction patterns showed that two Vibrio core group members, V. alginolyticus and V. parahaemolyticus were closely related having a similarity over 90%. Although the observed RFLP pattern for Grimontia hollisae shared several common bands with other Vibrio spp., G. hollisae results were still clearly distinct from Vibrio spp. RFLP types for all restriction enzymes tested. If restriction enzymes are aptly selected, PCR-RFLP analysis is still a rapid and effective tool for differentiating Vibrio species.

Characterization of Trichoderma spp. Associated with Green Mold of Oyster Mushroom by PCR-RFLP and Sequence Analysis of ITS Regions of rDNA

  • Park, Myung-Soo;Seo, Geon-Sik;Bae, Kyung-Sook;Yu, Seung-Hun
    • The Plant Pathology Journal
    • /
    • v.21 no.3
    • /
    • pp.229-236
    • /
    • 2005
  • Molecular profIles of PCR-RFLP and sequence analysis of internal transcribed spacer (ITS) regions of rDNA were compared between morphologically distinguishable species of Trichoderma isolated from substrates of oyster mushroom in Korea, T. atroviride, T. citrinoviride, T. harzianum, T. longibrachiatum, T. virens, and two unidentified species, Trichoderma sp. 1 and 2. PCR­RFLP analysis divided the Trichoderma spp. into six RFLP groups, A, B, C, D, E, and F. The RFLP groups were generally agreed with described morphological species, except that the RFLP group A containing the two unidentified species. A neighbor-joining tree based on ITS sequences well supported RFLP groups observed by RFLP analysis of ITS regions of rDNA. Additionally, the two unidentified species, Trichoderma sp. 1 and 2, which could not be distinguished by PCR­RFLP analysis, were separated in sequence analysis of ITS regions of rDNA.

T-RFLP Analysis of Microbial Community Structure in Leachate from Landfill Sites (폐기물매립장 침출수내 미생물군집 구조 해석을 위한 T-RFLP의 활용)

  • Yu, Jae-Cheul;Ishigaki, Tomonori;Kamagata, Yoichi;Lee, Tae-Ho
    • Journal of Korean Society of Environmental Engineers
    • /
    • v.32 no.4
    • /
    • pp.369-378
    • /
    • 2010
  • Microorganisms are key-role player for stabilization of landfill sites. In order to evaluate the availability of T-RFLP(Terminal Restriction Fragment Length Polymorphism) for monitoring microbial community variations during stabilization of landfill sites, the phylogenic diversity of microbial community in the leachate from 4 different full-scale landfills was characterized by T-RFLP based on bacterial 16S rDNA. Main population of microbial community analyzed by T-RFLP was significantly similar with that of microbial community analyzed by clone library analysis. The results of T-RFLP analysis for main population of microbial community in the leachate from landfills with different landfill structures, waste types and landfill ages showed apparently different microbial diversity and structures. Therefore, long-term monitoring of microbial community in leachate from landfill sites by using T-RFLP is expected to be available for evaluation of landfill stability.

Restriction Fragment Length Polymorphisms of Genomic DNA in Strains of Xanthomonas campestris pv. vesicatoria (지리적 기원이 다른 고추 더뎅이병균 균주 Genomic DNA의 RFLP 분석)

  • 정희정
    • Korean Journal Plant Pathology
    • /
    • v.12 no.2
    • /
    • pp.162-168
    • /
    • 1996
  • 우리 나라의 주요 고추 재배지와 미국, 대만, 호주, 아르헨티나에서 수집된 44 개 고추 더뎅이병균(Xanthomonas campestris pv. vesicatoria)균주간의 유전적변이를 genomic DNA의 restriction fragment length polymorphism(RFLP)에 의해 분석하였다. Genomic DNA RFLP profiles을 cluster 분석하여 얻은 dendrogram에서 지리적 기원이 다른 44개 균주들은 11개 RFLP 그룹으로 분류되었다. 외국 균주들은 genomic DNA의 RFLP 분석에 의해 모두 각각 다른 RFLP 그룹으로 분류되었다. 외국 균주들 중에서 미국 균주는 우리 나라 일부 균주들과 밀접한 유전적 관련성을 가지고 함께 cluster를 이루었는데, 이것은 이 균주들이 동일한 고추 더뎅이병균의 조상 균주 집단에서 유래했으리라는 것을 시사해 준다. 우리 나라 균주들은 6개의 RFLP 그룹으로 분류되었다. 대부분의 우리 나라 균주들은 가까운 cluster를 이루며 미국 균주를 제외한 외국 균주들과 뚜렷하게 구분되었다. 그러나 우리 나라 균주들 중에서 마산에서 수집된 Ms93-1은 다른 우리 나라 균주들과 뚜렷하게 구분되었다. 유전적으로 격리된 균주의 출현은 우리 나라에서 지리적 기원이 다른 고추 더뎅이병균 균주 사이에 이미 발생한 다양한 유전적 분화의 결과라고 추론된다.

  • PDF

Identification of Cryptosporidium in Environmental Sample using Nested PCR-RFLP and DNA Sequencing (Nested PCR-RFLP 및 DNA Sequencing을 이용한 환경시료에서의 크립토스포리디움 동정)

  • Park, Sangjung;Jeong, Hyanghee
    • Journal of Korean Society on Water Environment
    • /
    • v.24 no.6
    • /
    • pp.817-822
    • /
    • 2008
  • In order to identify various Cryptosporidium species in environment, nested PCR-RFLP and DNA sequencing method were used. The sensitivity of nested PCR-RFLP based on 18s rRNA gene was shown to 1 oocyst. Therefore, we applied nested PCR-RFLP method to environmental samples. As a result, only 4 samples out of 8 samples confirmed as Cryptosporidium parvum by standard method of Cryptosporidium were identified as Cryptosporidium parvum by nested PCR-RFLP and DNA sequencing method. The rest of 4 samples among 8 samples were identified as Cryptosporidium muris, Cryptosporidium bailey. Therefore, in addition to standard method of Cryptosporidium, supplementary verification through nested PCR-RFLP and DNA sequencing should be needed to give more accurate information about risk of Cryptosporidium.

Close relatedness of Acanthomoeba pintulosa with Accnthcmoebc palestinensis based on isoenzyme profiles and rDNA PCR-RFLP patterns (Acanthamoeba pustulosa와 A. palestinensis의 동위효소 및 rDNA PCR-RFLP 양상의 유사성)

  • 김영호;옥미선
    • The Korean Journal of Parasitology
    • /
    • v.34 no.4
    • /
    • pp.259-266
    • /
    • 1996
  • The taxonomic validity of morphological group III Accnthamoeba app. is uncertain. In the present study. six type strains of group III Aconthamoeba spry. , A. culbertsoni, A. heniyi, A. pustulosc, A. palestinensis, A. royrebn and A. lenticulnto were subjected for the evaluation or their taxonomic validity by comparison of the isoeneyme patterns by isoelectic focusing on polyacrylamide gels, mitochondrial DNA (Mt DNA) restriction fragment length polymorphism (RFLP) . and small subunit ribosomal DNA (ssu rDNA) PCR-RFLP patterns. The Mt DNA RFLP patterns were heterogeneous between the species. The type strains of A. pclestinensls and A. pustulosc showed almost identical patterns of isoenrymes and rDNA PCR-RFLP with an estimated sequence divergence of 2.6%. The other species showed heterogeneous patterns of isoenxymes and rDNA PCR- RFLP. It is likely that A. pustuLosc is closely related with A. palestinensis and that the former may be regarded as a junior synonym of the latter.

  • PDF

Effect of RFLP Marker of the Mitochondrial DNA D-Loop Region on Milk Production in Korean Cattle (한우 Mitochondrial DNA D-Loop 영역의 RFLP Marker가 산유량에 미치는 영향)

  • Chung Eui-Ryong;Chung Ku-Young
    • Food Science of Animal Resources
    • /
    • v.25 no.2
    • /
    • pp.218-225
    • /
    • 2005
  • This study was performed to determine sequence variation and RFLP of the mt DNA D-loop region using Southern blot hybridization analysis and to develop mt DNA marker affecting milk production traits in Hanwoo cows. The PCR was used to amplify an 1142 bp fragment within the D-loop region of mt DNA using specific primers. Mt DNA were digested with seven restriction enzymes and hybridized using DIG-labeled D-loop probe. The mt DNA RFLP polymorphisms were observed in the four enzymes, BamHI, RsaI, XbaI and HpaII. Nucleotide substitutions were detected at positions 441 (G/C), 469 (T/C), 503 (C/T), 569 (G/A), 614 (C/A) and 644 (C/T) of the mt DNA D-loop region between two selected lines. Significant relationship between the XbaI RFLP type and breeding value was found(p<0.05). Cows with A type had higher estimated breeding values than those with B type (P<0.05) between high and low milk production lines. Therefore, the RFLP marker of mt DNA could be used as a selection assisted tool for individuals with high milk producing ability in Hanwoo.

Incorporation of RAPD linkage Map Into RFLP Map in Glycine max (L, ) Merr (콩의 RAPD 연관지도를 RFLP 연관지도와 합병)

  • Choi, In-Soo;Kim, Yong-Chul
    • Journal of Life Science
    • /
    • v.13 no.3
    • /
    • pp.280-290
    • /
    • 2003
  • The incorporation of RAPD markers into the previous classical and RFLP genetic linkage maps will facilitate the generation of a detailed genetic map by compensating for the lack of one type of marker in the region of interest. The objective of this paper was to present features we observed when we associated RAPD map from an intraspecific cross of a Glycine max$\times$G. max, 'Essex'$\times$PI 437654 with the public RFLP map developed from an interspecific cross of G. max$\times$G. soja. Among 27 linkage groups of RAPD map, eight linkage groups contained probe/enzyme combination RFLP markers, which allowed us the incorporation of RAPD markers into the public RFLP map. Map position rearrangement was observed. In incorporating L.G.C-3 into the public RFLP linkage group a1 and a2, both pSAC3 and pA136 region, and pA170/EcoRV and pB170/HindIII region were in opposite order, respectively. And, pk400 was localized 1.8 cM from pA96-1 and 8.4 cM from pB172 in the public RFLP map, but was localized 9.9 cM from i locus and 18.9 cM from pA85 in our study. A noticeable expansion of the map distances in the intraspecific cross of Essex and PI 437654 was also observed. Map distance between probes pA890 and pK493 in L.G.C-1 was 48.6 cM, but it was only 13.3 cM in the public RFLP map. The distances from the probe pB32-2 to pA670 and from pA670 to pA668 in L.G. C-2 were 50.9 cM and 31.7 cM, but they were 35.9 cM and 13.5 cM in the public RFLP map. The detection of duplicate loci from the same probe that were mapped on the same or/and different linkage group was another feature we observed.

Genotyping of Six Pathogenic Vibrio Species Based on RFLP of 16S rDNAs for Rapid Identification

  • Yoon, Young-Jun;Im, Kyung-Hwan;Koh, Young-Hwan;Kim, Seong-Kon;Kim, Jung-Wan
    • Journal of Microbiology
    • /
    • v.41 no.4
    • /
    • pp.312-319
    • /
    • 2003
  • In an attempt to develop a method for rapid and accurate identification of six Vibrio species that are clinically important and most frequently detected in Korea, 16S rDNA restriction fragment length polymorphism (RFLP) of Vibrio type strains, as well as environmental isolates obtained from the Korean coastal area, was analyzed using ten restriction endonucleases. Digestion of the 16S rDNA fragments amplified by polymerase chain reaction (PCR) with the enzymes gave rise to 2~6 restriction patterns for each digestion for 47 Vibrio strains and isolates. An additional 2~3 restriction patterns were observed for five reference species, including Escherichia coli, Aeromonas hydrophila, A. salmonicida, Photobacterium phosphoreum, and Plesiomonas shigelloides. A genetic distance tree based on RFLP of the bacterial species correlated well with that based on 16S rDNA sequences. The very small 16S rDNA sequence difference (0.1%) between V. alginolyticus and V. parahaemolyticus was resolved clearly by RFLP with a genetic distance of more than 2%. RFLP variation within a species was also detected in the cases of V. parahaemolyticus, V. proteolyticus, and V. vulnificus. According to the RFLP analysis, six Vibrio and five reference species were assigned to 12 genotypes. Using three restriction endonucleases to analyze RFLP proved sufficient to identify the six pathogenic Vibrio species.

Identification of Fel ursi and Cattle and Pig Bile Juices by speciesspecific PCR and PCR-RFLP (종 특이 PCR과 PCR-RFLP를 이용한 웅담과 기타 담류의 감별 방법)

  • Kwon, Ki-Rok;Baek, Seung-Il;Choi, Suk-Ho
    • Journal of Pharmacopuncture
    • /
    • v.12 no.1
    • /
    • pp.13-20
    • /
    • 2009
  • Objective : This study developed species-specific PCR and PCR-RFLP to detect the adulteration of Fel ursi products with cattle and pig bile juices. Methods : All the primers for PCR and PCR-RFLP in this study were designed based on nucleotide sequences of cytochrome b genes in the mitochondria. Results : The species-specific PCR amplified a DNA fragment of 214, 214, 295, and 167 bp from Fel ursi product, bear fur, cattle bile juice, and pig bile juice, respectively. The survey using the speciesspecific PCR indicated that some of commercial Fel ursi products were adulterated with cattle and pig bile juices. PCR-RFLP using the restriction endonucleases, HaeIII and HinfI enabled differentiation among Fel ursi product, cattle bile juice, and pig bile juice. Bear furs from two animals showed variations in PCR-RFLP patterns with HaeIII. Discussion : The detection methods of the species-specific PCR and PCR-RFLP could be useful in eliminating adulterated Fel ursi products from the market.