• Title/Summary/Keyword: RAPD-PCR

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Antibiotic Susceptibility Pattern and Molecular Typing By PCR-RAPD Analysis of Clinical and Environmental Isolates of Pseudomonas aeruginosa

  • Oluborode, O.B.;Smith, S.I.;Seriki, T.A.;Fowora, M.;Ajayi, A.;Coker, A.O.
    • Microbiology and Biotechnology Letters
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    • v.46 no.4
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    • pp.434-437
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    • 2018
  • Pseudomonas aeruginosa accounts for a significant proportion of nosocomial infections. This study examined the antimicrobial susceptibility pattern and clonal relatedness of P. aeruginosa isolates of clinical and environmental origin. These isolates displayed susceptibility to levofloxacin, ciprofloxacin, gentamicin, imipenem, and ceftazidime of 65.0%, 62.5%, 90.0%, 100%, and 85%, respectively. PCR-RAPD analysis of the P. aeruginosa isolates revealed marked variation. No correlation was observed between the antibiotic resistance profiles and the DNA typing patterns.

Studied on Amplificative Efficiency of PCR of Predigested template DNA and GC Contents for RAPD Analysis in the Silkworm, Bombyx mori (누에의 RAPD 분석을 위한 primer의 GC 함량과 사전 제한효소 처리한 주형 DNA의 PCR 증폭효율에 관한 연구)

  • 이진성;황재삼;이상몽;황석조;강현아;성승현;서동상
    • Korean journal of applied entomology
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    • v.35 no.1
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    • pp.58-65
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    • 1996
  • This experiment was carried out to evaluate the effect of sublethal doses of BPMC, etofenprox, and buprofezin on N. lugens. and its predator C. lividipennis. Buprofezin was found to be the most toxic to N. lugens and the most safe to C. lividipennis among the three insecticides, based on LD50 values. Selective toxicity index calculated by dividing LDSo value of C. lividipennis by that of N. lugens indicated that buprofezin was very safe to C. lividipennis, showing selective toxicity of 2703.3. Longevity and fecundity of N. lugens treated with LDIU and LDm of buprofezin and BPMC were not significantly different with those of untreated brown planthoppers. However, egg hatchability' of N. lugens was greatly reduced when treated with LDm of buprofezin, having the highest inhibition rate of 17.7%. Hatchability of eggs from insects treated with BPMC was similar to that of control. The oviposited peak of treated hoppers appeared late as compared to the untreated which showed the peak at early part of the ovipositional period. The longevity and fecundity of C. lividipennis treated with BPMC were significantly reduced as compared with the untreated. Etofenprox also induced fecundity reduction when treated with LDlo, and LDm. However, C. lividipennis treated with sublethal doses of buprofezin showed no redution in logevity and fecundity. From these results, it may be said that buprofezin can be used to control brown planthopper without disrupting of C. lividipennis population in the rice field.

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Absence of DNA Polymorphisms in Myzus persicae (Homoptera: Aphididae) in Relation to their Host Plants (기주식물 종류에 따른 복숭아혹진딧물(Myzus persicae)의 DNA Polymorphism 비교)

  • H. J. Kim;K. S. Boo;K. H. Cho
    • Korean journal of applied entomology
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    • v.35 no.3
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    • pp.209-215
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    • 1996
  • DNA polymorphisms were analyzed for 8 clones of the green peach aphid, Myzus persicae Sulzer, by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). The insect has different host preferences and was even classified into two different species, M. persicae Sulzer and Myzus nicotinae Blackman by their morphological characters, but this point is still in arguement. To identify the differences between two types of the green peach aphid by RAPD-PCR, the template DNA was extracted from 4 clones each of tobacco-feeding and non-tobacco-feeding forms and one hundred primers of 10-nucleotideslong were tested in PCR. The amplified DNAs were analyzed by agarose gel electrophoresis. Eighty-three primers gave amplified DNA fragments with 1 to 22 in number and 500 to 20,000 base pairs in length, but no amplification was observed in the other 17 primers. The average number of fragment per each amplification was about 13. In the case of 82 out of 83 random primers, band patterns of amplified DNA were identical among 8 clones, even though some differences were noticed in the intensity of specific bands. Polymorphism was detected by only one primer within the tobacco-feeding forms, but not between the two host types. The results did not detect any relationship between RAPD polymorphism and their host preference.

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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.

Development of RAPD-SCAR and RAPD-generated PCRRFLP Markers for Identification of Four Anguilla eel Species

  • Kim, Woo-Jin;Kong, Hee-Jeong;Kim, Young-Ok;Nam, Bo-Hye;Kim, Kyung-Kil
    • Animal cells and systems
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    • v.13 no.2
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    • pp.179-186
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    • 2009
  • Discriminating between eel species of the genus Anguilla using morphological characteristics can be problematic, particularly in the glass eel and elver stages. In this study, sequence-characterized amplified region (SCAR) and polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) markers were developed for the identification of Anguilla japoniea, Anguilla btcoior bicaor. Anguilla rostrata, and Anguilla anguilla. Random amplified polymorphic DNA (RAPD) fragments from A. japoniea (362 bp), A. bicolor bicctor (375 bp), A. rostrata (375 bp), and A. anguilla (375 bp) were isolated, sequenced, and converted to SCAR markers. The principal difference between the SCARs of A. japoniea and the three other species is the absence of a 13 bp deletion in the A. japoniea SCAR. Specific PCR primers amplified a 290 bp fragment for A. japoniea and 303 bp fragments for A. bicolor bicoior. A. rostrata, and A. anguilla. Restriction enzyme digestion with Taql, Mael, and Tru9l yielded PCR-RFLP patterns with differences that, when analyzed together, are sufficient for distinguishing each of the four eel species. In addition, RAPD fragments for A. japoniea (577 bp), A. bicoior bicoor (540 bp), A. rostrata (540 bp), and A. anguilla (509 bp) were also isolated and sequenced. The A. japoniea, A. bicoior blcoior. A. rostrata, and A. anguilla PCR products contain ten, nine, nine, and eight tandem repeats, respectively, of a 37 bp sequence. These results suggest that SCAR and PCR-RFLP markers and repeat numbers for specific loci will be useful for the identification of these four Anguilla eel species.

RAPD Analysis of DNA Polymorphism and Genetic Species-Specificity Using PCR Technique in the Marine Microalgae

  • Yoon, Jong-Man;Chang, Kye-Nam
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.487-488
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    • 2000
  • Genomic DNA was isolated from the marine microalgae representing genetic characteristics and genomic polymorphisms by polymerase chain reaction amplification of DNA as arbitrary primers. The electrophoretc analysis of PCR-RAPD products showed hig levels of variation between different genus and little variation between different species. Outer of these primers, 6 generated 248 highly reproducible RAPD markers, producing almost seven polymorphic bands per primers. The degree of similarity frequency between Chaetoceros gracilis and Chaetoceros calcitrans species showed 90% as calculated by sharing analysis. The RAPD polymorphism generated by this primer may be used as a genetic marker for genus or species identification in important marine microalgae. (omitted)

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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.

Relationship of Lycoris (Amaryllidaceae) Based on RAPD Markers (RAPD markers에 의한 상사화속 식물의 유연관계)

  • Tae, kyoung-Hwan;Kim, Yong Hyun;Shin, Young-Hwa;Kang, Shin-Ho;Kim, Joo-Hwan;Ko, Sung-Chul
    • Korean Journal of Plant Taxonomy
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    • v.38 no.1
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    • pp.17-29
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    • 2008
  • Phylogenetic relationships were examined for 17 taxa of Lycoris by RAPD analysis. The length of the amplified DNA fragments ranged from 300 bp to 1,700 bp. 57 scorable RAPD markers were observed from PCR reactions with five random oligoprimers. The analysis by UPGMA sepatated the examined taxa of Lycoris into were clusters. First group was comprised of ten taxa of L. chinensis var. sinuolata, L. sanguinea var. koreana, L. uydoensis, L. flavescens, L. radiata var. pumila, L. radiata, L. squamigera, L. chejuensis, L. aurea and L. guangxiensis, second group of L. haywardii, L. sprengeri, L. rosea, L. straminea and L. houdyshii, third group of outgroup of Narcissus tazetta var. chinensis and Crinum asiaticum var. japonicum. From the viewpoint of cytological characters such as polyploidy and karyotype, the RAPD analysis was very useful to show the relationship among the intraspecific taxa of Lycoris.

Linkage Analysis of both RAPD and I-SSR Markers using Haploid Genome from a Single Tree of Pinus densiflora S. et Z. (소나무 단일(單一) 모수(母樹)의 반수체(半數體) 게놈을 이용(利用)한 RAPD 및 I-SSR 표식자(標識子)의 연관분석(連關分析))

  • Hong, Yong-Pyo;Chung, Jae-Min;Kim, Yong-Yul;Jang, Suk-Seong
    • Journal of Korean Society of Forest Science
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    • v.89 no.4
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    • pp.536-542
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    • 2000
  • A linkage map for Japanese red pine (Pinus densiflora) was constructed on the basis of two DNA marker systems of random amplified polymorphic DNAs (RAPDs) and inter-simple sequence repeats (I-SSR). Haploid genomic DNAs were extracted from megagametophyte tissues of 96 individual seeds in a single tree. A total of 98 DNA markers including 52 RAPD markers amplified by 25 primers and 46 I-SSR markers amplified by 18 primers were verified as Mendelian loci showing 1 : 1 segregation in 96 megagametophytes which were ${\chi}^2$-tested at 5% significance level. Of them, 63 segregating loci turned out to be linked into 20 linkage groups by the two-point analysis. However, 35 loci (17 RAPD and 18 I-SSR) of the 98 segregating loci did not coalesced into any linkage groups at a LOD of 3.0. The linked 63 loci were separated by an average distance of about 25.5 cM, which were spanned 1097.8 cM as a whole. The minimum and maximum map distances of the linkage groups were 4.3 cM and 54.9 cM, respectively. Incorporation of I-SSR loi into linkage map of RAPD loci resulted in extended and partially more saturated linkage blocks.

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Parameters Affecting Polymerase Chain Reaction in RAPD Analysis of Pleurotus spp. (느타리버섯속(屬)의 DNA 다형성분석(多型性分析)에 영향(影響)을 미치는 PCR 조건(條件))

  • Kim, Beom-Gi;Jeong, Mi-Jeong;Lee, Chang-Soo;Lee, Hee-Kyung;Yoo, Young-Bok;Ryu, Jin-Chang
    • The Korean Journal of Mycology
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    • v.23 no.3 s.74
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    • pp.202-208
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    • 1995
  • This study describes the effects of several components on PCR amplification used for RAPD. We used different concentrations of reaction components to obtaine discrete and reproducible PCR products from Pleurotus cornucopiae. The optimum concentrations of reaction components were found to be 80 ng of template DNA, 30 pmole of 10-mer primer, $200\;{\mu}M$ dNTP, 2mM $MgCl_2$, 50 mM KCl, 10 mM Tris-HCl(pH 9.0), 0.1% Triton X-100, 1.5 unit of Taq DNA polymerase (promega) in $50\;{\mu}l$ reaction volume. The optimum annealing temperature was $35^{\circ}C$. These results proved to be valuable for characterization of Pleurotus spp.

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