• Title/Summary/Keyword: RAPD fingerprinting

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Enhanced Discrimination of Listeria spp. Using RAPD Fingerprinting Complemented by Ribotyping-PCR (리스테리아균의 특성분석을 위한 Molecular Typing 방법의 상호보완)

  • 임형근;홍종해;박경진;최원상
    • Journal of Life Science
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    • v.13 no.5
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    • pp.699-704
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    • 2003
  • The results typed by random amplification of polymorphic DNA (RAPD) were compared with those obtained by Enterobacterial repititive intergenic consensus (ERIC) fingerprinting and ribotyping-PCR. The discriminatory power of RAPD typing was the best among the methods tested. RAPD typing with two different primers for 13 Listeria spp. reference strains produced 11 patterns each. In contrast, ERIC fingerprinting produced 9 patterns and ribotyping-PCR produced 7 patterns each. Composite of two separate RAPD (Lis 11 and primer 6) results or RAPD (Lis11)/ ribotyping-PCR differentiated all 13 Listeria spp. reference strains. Therefore, composite of 2 separate RAPD (Lis11 and primer 6) or composite of RAPD (Lis11)/ribotyping-PCR is expected the most promising approach for typing field isolated Listeria spp. strains.

RAPD Fingerprinting for the Species Identification of Animals

  • Huang, Mu-Chiou;Horng, Yan-Ming;Huang, Hsiu-Lin;Sin, Yen-Long;Chen, Ming-Jaw
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.10
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    • pp.1406-1410
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    • 2003
  • The studies were based on the RAPD fingerprinting for the species identification of animals. The genomic DNA samples of ostriches, Taiwan local chickens, Aboracres broilers, Leghorn chickens, quails, doves, emus, Beltville small white turkeys, pheasants, Chinese geese, mule ducks, Holstein cattle and Landrace pigs were amplified with random primers by RAPD-PCR for fingerprinting. The results showed that the varied band patterns of DNA fingerprints were generated from templates depending on the kinds of primers or animal species. The same primer applied to the same breed, all of the main bands are similar, but which were different among species. In order to try to identify the species from the mixture of meat by RAPD fingerprinting, the meat of ostrich and cattle was mixed in different ratios for this study. The results showed that it could be easily and precisely distinguished according to the band distribution of RAPD patterns.

Differentiation of Salmonella typhimurium from Gram-negative Intestinal Microbes by Randomly Amplified Polymorphic DNA (RAPD) Fingerprinting

  • Jin, Un-Ho;Chung, Tae-Wook;Kim, June-Ki;Nam, Kyung-Soo;Ha, Sang-Do;Kim, Cheorl-Ho
    • Journal of Microbiology
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    • v.38 no.1
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    • pp.8-10
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    • 2000
  • In order to rapidly identify and differentiate Salmonella typhimurium from the intestinal gram-negative bacteria, randomly amplified polymorphic DNA (RAPD) fingerprinting of Salmonella typhimurium was carried out using random primers designated OPA-13 (5'-CAGCACCCAC-3'), OPB-10 (5'-CGTCTGGGAC-3'), OPB-18 (5'-CCACAGCAGT-3'), and OPJ-10 (5'-AAGCCCGAGG-3'), and its patterns compared with 6 representive intestinal, gram-negative bacterial strains, Vibrio parahaemolyticus, V. vulnificus, Enterobacter cloacae, Escherichia coli O157:H7, Pseudomonas aeruginosa, and Proteus sp., which are often found in foods. S. typhimurium had unique and distinct fingerprinting patterns. RAPD fingerprinting is thus concluded to be a rapid and sensitive method for the identification of S. typhimurium compared to conventional culturing procedures or immunoassays.

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Genetic Diversity Estimation of the Rice Mutant Lines Induced by Sodium Azide

  • Shin, Young-Seop;Jeung, Ji-Ung
    • Korean Journal of Breeding Science
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    • v.43 no.1
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    • pp.23-31
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    • 2011
  • To investigate dose-effect of a chemical mutagen, sodium azide on a rice elite line, Suweon472, seed aliquots were treated with five different concentrations of sodium azide. The degree of mutation levels of each aizde concentration were estimated by using DNA fingerprinting techniques such as RAPD and AFLP. Some selected mutant lines ($M_4$) were also subjected for DNA fingerprinting to estimate their mutation levels by comparing the banding patterns of the wild type, Suweon 472. RAPD and AFLP fingerprinting patterns indicated that dose-effect of different azide concentrations was not clear. With allele description of detected AFLPs among favorable mutant lines, it was possible to discriminate each mutant line from others which have similar phenotypes and reactions against pathogens. AFLP fingerprinting patterns of waxy mutant lines, otherwise, were highly homogeneous as well as their phenotypic and agronomic characters.

DNA Fingerprinting of Rice Cultivars using AFLP and RAPD Markers

  • Cho, Young-Chan;Shin, Young-Seop;Ahn, Sang-Nag;Gleen B. Gregorio;Kang, Kyong-Ho;Darshan Brar;Moon, Huhn-Pal
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.44 no.1
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    • pp.26-31
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    • 1999
  • This experiment was conducted to evaluate genetic variation in 48 rice accessions (Oryza sativa L.) using AFLP and RAPD markers. For AFLP, a total of 928 bands were generated with 11 primer combinations and 327 bands (35.2%) of them were polymorphic among 48 accessions. In RAPD analyses using 22 random primers 145 bands were produced, and 121 (83.4%) were polymorphic among 48 accessions. Each accession revealed a distinct fingerprint by two DNA marker systems. Cluster analysis using AFLP-based genetic similarity tended to classify rice cultivars into different groups corresponding to their varietal types and breeding pedigrees, but not using RAPD-based genetic similarity. The AFLP marker system was more sensitive than RAPD in fingerprinting of rice cultivars with narrow genetic diversity.

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Genomic Fingerprinting Patterns of Bifidobacteria Isolated from Healthy Koreans Using ERIC-, TAP-, and BOX-PCR (건강한 한국인으로부터 분리된 비피도박테리아의 ERIC-, TAP-, BOX- 중합효소연쇄반응을 이용한 유전자 지문 분석)

  • Lee, Do-Kyung;Kang, Byung-Yong;Chung, Myung-Jun;Lee, Kang-Oh;Kim, Kyung-Jae;Ha, Nam-Joo
    • Environmental Analysis Health and Toxicology
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    • v.23 no.1
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    • pp.1-9
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    • 2008
  • 유산균인 비피도박테리아는 사람과 동물에서 유익한 프로바이오틱 미생물로 알려져 있다. 본 연구에서는 이러한 비피도박테리아 균주의 분류를 위한 repetitive DNA element PCR fingerprinting (ERIC-또는 TAP-PCR)의 사용을 평가하였다. 사람분변으로부터 분리한 알려지지 않은 비피도박테리움 균주와 한국생명공학연구원 생물자원센터로부터 분양받은 표준균주를 가지고 분류 및 동정에 ERIC-PCR과 TAP-PCR을 이용한 RAPD-fingerprinting을 수행하였다. 그 결과 비피도박테리움 균주에 대한 속과 종단위의 분류가 가능하였으며, 실험에 사용된 모든 비피도박테리움 균주는 RAPD-fingerprinting 분석을 통해 유전적 다양성을 확인하였다. 또한 ERIC2와 TAP1 프라이머를 이용한 실험에서는 Bifidobacterium adolescenits 특이 유전자 단편을 확인하였으며 이는 B. adolescenits 균주의 동정에 유용할 것으로 사료된다.

Efficiency of RAPD and ISSR Markers in Differentiation of Homo- and Heterokaryotic Protoclones of Agaricus bisporus

  • Mahmudul, Islam Nazrul;Bian, Yin-Bing
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.683-692
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    • 2010
  • Morphologically, nine different slow-growing protoclones were screened from regenerated protoplasts of heterokaryotic Agaricus bisporus. As such, the present study is the first report on differentiating homo- and heterokaryotic protoclones using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers. Among 80 primers tested, the seven ISSR and seven RAPD primers selected for the analysis generated a total of 94 ISSR and 52 RAPD fragments, respectively. The ISSR fingerprinting also detected more polymorphic loci (38.29%) than the RAPD fingerprinting (34.61%). A principal coordinate analysis (PCA) was employed to evaluate the resolving power of the markers as regards differentiating protoclones. As a result, the mean polymorphism information content (PIC) for each marker system (i.e., 0.787 for RAPD and 0.916 for ISSR) suggested that ISSR is more effective for determining polymorphisms. The dendrograms constructed using RAPD, ISSR, and an integrated RAPD and ISSR marker system were highly correlated with one another as revealed by a high Mantel correlation (r= 0.98). The pairwise similarity index values also ranged from 0.64 to 0.95 (RAPD), 0.67 to 0.98 (ISSR), and 0.67 to 0.98 (RAPD and ISSR), whereas the mean similarity index values of 0.82, 0.81, and 0.84 were obtained for the RAPD, ISSR, and combined data, respectively. As there was a good correspondence between the RAPD and ISSR similarity matrices, ISSR would appear to be an effective alternative to RAPD in the genetic diversity assessment and accurate differentiation of homo- and heterokaryotic protoclones of A. bisporus.

Toxin Gene Typing, DNA Fingerprinting, and Antibiogram of Clostridium perfringens Isolated from Livestock Products

  • Lee, Seung-Bae;Choi, Suk-Ho
    • Food Science of Animal Resources
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    • v.26 no.3
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    • pp.394-401
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    • 2006
  • Forty Clostridium perfringens isolates were obtained from twelve animal products, following the examination of eighty six beef, pork, broiler chicken and salami meat products, and eleven milk powder products. There were 21 isolates from salami stored at $25^{\circ}C$, 3 isolates from pork, 4 isolates from beef, 9 isolates from broiler chicken, and 3 isolates from milk powder. Only the cpa gene encoding a toxin among the 5 toxin genes tested (cpa, cpb, etx, iap, and cpe) was detected in all forty isolates, suggesting contamination with C. perfringens type A. DNA fingerprinting analysis using PCR of the tRNA intergenic spacer (tDNA-PCR) and the 16S-23S internal transcribed spacer (ITS-PCR), and randomly amplified polymorphic DNA (RAPD) analysis were attempted to differentiate the isolates. RAPD analysis was the most discriminating method among the three PCR analyses. Isolates from the same products tended to show similar RAPD patterns. Antimicrobial susceptibility tests showed that some isolates from broiler chickens had the same antibiogram with multiple resistance to streptomycin, colistin, and ciprofloxacin. Antibiograms were similar between isolates from the same livestock products, but differed considerably between the products.

Molecular Typing of Leuconostoc citreum Strains Isolated from Korean Fermented Foods Using a Random Amplified Polymorphic DNA Marker

  • Kaur, Jasmine;Lee, Sulhee;Sharma, Anshul;Park, Young-Seo
    • Food Engineering Progress
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    • v.21 no.2
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    • pp.174-179
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    • 2017
  • For preliminary molecular typing, PCR-based fingerprinting using random amplified polymorphic DNA (RAPD) is the method of choice. In this study, 14 bacterial strains were isolated from different Korean food sources, identified using 16S rRNA gene sequencing, and characterized through RAPD-PCR. Two PCR primers (239 and KAY3) generated a total of 130 RAPD bands, 14 distinct PCR profiles, 10 polymorphic bands, one monomorphic band, and four unique bands. Dendrogram-based analysis with primer 239 showed that all 14 strains could be divided into seven clades out of which clade VII had the maximum of seven. In contrast, dendrogram analysis with the primer KAY3 divided the 14 L. citreum strains into four clades out of which clade IV consisted of a maximum of 10 strains out of 14. This research identified and characterized bacterial populations associated with different Korean foods. The proposed RAPD-PCR method, based on sequence amplification, could easily identify and discriminate the lactic acid bacteria species at the strain-specific level and could be used as a highly reliable genomic fingerprinting tool.

Molecular Typing of Pseudomonas aeruginosa by Randomly Amplified Polymorphic DNA

  • Byoung-Seon Yang
    • Biomedical Science Letters
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    • v.9 no.4
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    • pp.183-187
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    • 2003
  • Pseudomonas aerugionsa is a commonly isolated nosocomial pathogen. DNA fingerprinting of P. aerugionsa is examined by randomly amplified polymorphic DNA (RAPD). In this study, P. aeruginosa were isolated from environmental and clinical specimens and the molecular typing of the microorganisms was investigated by RAPD. Thirty strains of P. aeruginosa were selected from the strains isolated formerly and submitted for type identification to the University Hospital. 15 strains of P. aeruginosa were received from Chungnam University Hospital and 14 strains from Gyeongsang University Hospital. DNA of P. aeruginosa was extracted by Qiagen genomic DNA kit. PCR mixtures were set up and incubated, Reactions mixtures were made to be optimal for P. aeruginosa. RAPD typing analysis was carried out by the multivariate statistical program (MVSP) V3.0. RAPD type I was the most common pattern and included 23 strains. Most of strains from Gyeongsang University Hospital belonged to RAPD type lb and 15 strains from Chungnam University Hospital to RAPD type I or II. RAPD typing of P. aeruginosa isolated from the environmental and clinical specimens was very simple and reproducible.

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