• Title/Summary/Keyword: DNA fingerprinting

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DNA Fingerprinting of Jute Germplasm by RAPD

  • Hossain, Mohammad Belayat;Haque, Samiul;Khan, Haseena
    • BMB Reports
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    • v.35 no.4
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    • pp.414-419
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    • 2002
  • The genotype characteristic of cultivars was investigated, along with varieties of both of the jute species, Corchorus olitorius and Corchorus capsularis, in the germplasm collection at the Bangladesh Jute Research Institute (BJRI). DNA fingerprinting was generated for 9 different varieties and 12 accessions of jute cultivars by using random amplified polymorphic DNA(RAPD). A total of 29 arbitrary oligonucleotide primers were screened. Seven primers gave polymorphism within the varieties, and 6 primers detected polymorphism within the accessions that were tested. A dendrogram was engendered from these data, and this gave a distinct clustering of the cultivated species of jute. Therefore, we generated RAPD markers, which are species-specific. These primers can distinguish between C. olitorius and C. capsularis. From the dendrogram that we generated between the various members of these two species, we found the existing genetic classification that agrees with our molecular marking data. A different dendrogram showed that jute accessions could be clustered into three groups. These data will be invaluable in the conservation and utilization of the genetic pool in the germplasm collection.

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 Characterization of Small-Spored Alternaria Species (소형의 포자를 형성하는 Alternaria 균류의 분자생물학적 특징)

  • Kim, Byung-Ryun;Park, Myung-Soo;Cho, Hye-Sun;Yu, Seung-Hun
    • Research in Plant Disease
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    • v.11 no.1
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    • pp.56-65
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    • 2005
  • To establish taxonomic system of morphologically similar species of small-spored Alternaria, phylogenetic analysis of internal transcribed spacer (ITS 1, ITS 2 and 5.8S rDNA) and mitochondrial small subunit (mt SSU) rDNA sequences and URP-PCR fingerprinting analysis from 11 species ofAlternaria were performed. Phylogenetic analysis of ITS and mt SSU rDNA sequences revealed that 10 out of 11 species of the smallspored Alternaria were phylogenetically identical with a bootstrap value of 100%. A. infectoria only was phylogenetically differentiated from the other species. The results suggest that the 10 small-spored Alternaria species are very closely related evolutionally and the markers can not be used for differentiation of the smallspored Alternaria species. URP-PCR fingerprinting analysis from eleven species of smallspored Alternaria using 10 URP primers showed that it was possible to differentiate the species, although genetic similarities were found among the species. The Alternaria sp. from common pokeweed could be distinguished from other species by URP-PCR analysis, and it was considered as a new species. A. infectoria could be easily distinguished from the other 10 species by phylogenetic analysis of ITS and mt SSU rDNA sequences and the URPPCR fingerprinting analysis.

Application of rDNA-PCR Amplification and DGGE Fingerprinting for Detection of Microbial Diversity in a Malaysian Crude Oil

  • Liew, Pauline Woan Ying;Jong, Bor Chyan
    • Journal of Microbiology and Biotechnology
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    • v.18 no.5
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    • pp.815-820
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    • 2008
  • Two culture-independent methods, namely ribosomal DNA libraries and denaturing gradient gel electrophoresis (DGGE), were adopted to examine the microbial community of a Malaysian light crude oil. In this study, both 16S and 18S rDNAs were PCR-amplified from bulk DNA of crude oil samples, cloned, and sequenced. Analyses of restriction fragment length polymorphism (RFLP) and phylogenetics clustered the 16S and 18S rDNA sequences into seven and six groups, respectively. The ribosomal DNA sequences obtained showed sequence similarity between 90 to 100% to those available in the GenBank database. The closest relatives documented for the 16S rDNAs include member species of Thermoincola and Rhodopseudomonas, whereas the closest fungal relatives include Acremonium, Ceriporiopsis, Xeromyces, Lecythophora, and Candida. Others were affiliated to uncultured bacteria and uncultured ascomycete. The 16S rDNA library demonstrated predomination by a single uncultured bacterial type by >80% relative abundance. The predomination was confirmed by DGGE analysis.

Differentiation Among Commercial Strains of Pleurotus spp. Based on DNA Fingerprinting Using Universal Rice Primer (URP) (Universal Rice Primer(URP)에 의한 DNA 핵산지문법을 이용한 느타리의 유통 품종간 구분)

  • Seo, Kyoung-In;Jang, Kab-Yeul;Yoo, Young-Bok;Park, Soon-Young;Kim, Kwang-Ho;Kong, Won-Sik
    • The Korean Journal of Mycology
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    • v.36 no.2
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    • pp.130-137
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    • 2008
  • To distinct the commercial strains in Pleurotus spp., 81 strains in eight Pleurotus species were used. DNA fingerprinting using URP-PCR was conducted to determine the phylogenetic relationships among Pleurotus strains. DNA profiles of Pleurotus species obtained by twelve URP primers were analyzed for genetic similarity by NTSYS program. We could divide strains into ten clusters, in which three of them belong to P. ostreatus and the others to the different species, respectively. At the 76% similarity level, 70 P. ostreatus strains were distinguished into three clusters. Cluster I contained 35 strains and some of them showed almost 100% similarity, one strain closely related to Weonhyeong and six strains closely related to Wangheukpyeong. In cluster II, twenty-one out of 23 strains showed 100% to Suhan. Cluster III contained twelve strains, including six strains closely related to Chunchu-2. The results suggested that there are many same strains with different names in mushroom spawn market.

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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    • v.9 no.3
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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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DNA fingerprinting analysis of maize varieties and parental lines using microsatellite markers (Microsatellite 마커를 이용한 옥수수 품종 및 자식 계통에 대한 DNA Fingerprinting 분석)

  • Kwon, Yong-Sham
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.367-375
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    • 2016
  • In the present study, we conducted genetic characterization of 90 commercial maize varieties and parental lines using microsatellite markers. Thirteen microsatellite markers were selected from 100 primer pairs in the maize genome data on the basis of polymorphism information contents (PIC) value and distinct amplification products. These markers detected 5 to 24 alleles, with an average of 13.69. The mean PIC value was 0.865 and ranged from 0.716 to 0.942. The unweighted pair-group method with arithmetical average (UPGMA) analysis was conducted for constructing the dendrogram using Jaccard's genetic similarity coefficient. The genetic similarity varied from 0.07 to 0.824. Thirteen microsatellite markers identified all 90 maize varieties and parental lines. The maize varieties were clustered into 5 major groups consistent with type and pedigree information. The microsatellite profile database of maize varieties could be used to select comparative varieties through genetic relationship analysis between existing varieties and candidate varieties in distinctness tests.

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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