• Title/Summary/Keyword: RAPD method

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Random Amplified Polymorphic DNA (RAPD) Analysis for Extended Spectrum ${\beta}-Lactamase$ Producing Klebsiella pneumonia Isolated from Clinical Specimens in Korea

  • Kim Yun-Tae
    • Biomedical Science Letters
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    • v.12 no.3
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    • pp.267-274
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    • 2006
  • Klebsiella pneumoniae is the leading cause of nasocomial infection and the most commonly isolated from clinical specimens. $Extended-spectrum-{\beta}-lactamase$ (ESBL) producing K. pneumoniae infection was associated with a significantly longer duration of hospital stay and greater hospital charges. The purpose of this study is to investigate the antibiotic resistant patterns and the DNA fingerprint types of extended-spectrum ${\beta}-lactamase$ (ESBL) producing K. pneumoniae. 223K. pneumoniae strains were collected from three general hospitals with more than 500 beds in Busan, Korea from September 2004 to October 2005. The minimum inhibitory concentration (MIC) of antibiotics was measured using the Gram negative susceptibility (GNS) cards of VITEK (Vitek system, Hazelwood Inc., MO). Random amplified polymorphic DNA method was used to detect DNA fingerprint of the organisms. Of the 226 K. pneumoniae isolates 65 ESBL-producing K. pneumoniae strains were detected by the Vitek system and confirmed by the double-disk synergy test. All the 65K. pneumoniae strains were resistant cefazolin, cefepime, ceftriaxone and aztreonam, and 83.0% of the organisms were resistant to ampicillin/sulbactam, 66.1% to tobramycin, 67.6% to piperacillin/tazobactam, 61.5% to ciprofloxacin, and 47.6% to trimethoprim/sulfamethoxazole and 43.0% to gentamicin. The RAPD patterns were distincted as 10 types by three random 10-mer primers (208, 272, 277). Among ten type patterns, three types (Ic, IIb, IIIe) were remarkably represented at patient of internal department, nerve surgery department, general surgery department, and neonatal room. These results indicate that RAPD can be useful for DNA of strains typing in the epidemiological investigations. Therefore more investigation are needed in order to prevent the ESBL type-producing K. pneumoniae from spreading resistance to oxyimino cephalosphorin antibiotics.

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Morphological and Genetic Diversity of Korean Native and Introduced Safflower Germplasm

  • Shim Kang-Bo;Bae Seok-Bok;Lim Si-Kyu;Suh Duck-Yong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.4
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    • pp.337-341
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    • 2004
  • Morphological and genetic diversity of thirty nine safflower germplasm were collected and evaluated by Principal Component Analysis (PCA) and Random Amplified Polymorphic DNA (RAPD) method. Stem length and seeding to flowering days of the safflower germplasm showed $26\~117cm\;and\;76\~179$ days of variation respectively. USA originated germplasm showed higher oil content as $39\%$, but that of Japanese showed lower as $26\%$. PCA made three different cluster groups according to some agronomic characteristics of safflower. Korea originated germplasm showed similar cluster group with that of collected from USA in the PCA of stem length. But in the seeding to flowering days, it showed similar cluster pattern with that of collected from Japan rather than USA. In the experiment of RAPD analysis, total five primers showed polymorphism at the several chromosomal loci. Korea, China Japan and South Central Asia originated germplasm were differently classified with USA and South West Asia originated germplasm with lower similarity coefficient value (0.47). Most of Korea originated germplasm were grouped with South Central Asia originated germplasm with higher similarity coefficient value (0.74) conferring similar genetic background between both of them. China and Japan originated germplasm were dendrogramed with Korea originated germplasm at the 0.65 and 0.50 similarity coefficient values respectively. Some common results were expected from both of PCA and RAPD analysis, but lower genetic heritability caused by relative higher portion of environmental variance and environment by genotype interaction at the expression of those of agronomic characteristics made constraint to find any reliable results.

Systematic Relationships among Species of the Genus Dendronephthya (Alcyonacea: Octocorallia; Anthozoa) Based on RAPD Analysis

  • Song, Jun-Im;Lee, Young-Ja
    • Animal cells and systems
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    • v.4 no.1
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    • pp.1-7
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    • 2000
  • The genus Dendronephthya, generally known as soft corals, is reported as an abundant and variable taxon. They mostly distribute in warmer waters of the Undo-Pacific Ocean region including Korea. In spite of their abundance and ecological importance as habitats of marine organisms, there are difficulties in the study of their identification and systematics because they have morphological variabilities and limited taxonomec characters. To resolve the problems, we attempted to elucidate the genetic relationships in the genus Dendronephthya by using random amplified polymorphic DNA (RAPD) analysis. This study was based on eight dendronephthian species and one Alcyoniidae species, Alcyonium gracillimum, as an outgroup. The results from all analysis suggest that they could be classified into four groups by the growth form and the anthocodial grades as follows: the first one,D. putteri and D. suensoni with the divaricate form and VI grade; the second one,D. sp.1 and D. sp.2 with the divaricate form and III or IV grade; the third one, D. gigantea and D. aurea being closer than D. spinifera with the glomerate form and III grade; the last one, D. castanea related to D. gigantea rather than D. putteri with the umbellate form and IV grade. Moreover, the divaricate form was separated from the group of the glomerate and umbellate form. At the intraspecies level, the types of the D. castanea, D. gigantea and D. spinifera were separated depending on the feature of spicules in the polyp head, and the coloration could not influence genetic variation. From this study, we can confirm that their morphological characters are compatible with the genetic variation, also RAPD analysis is a very useful method for resolving the systematic relationships of den-deonephthians.

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Analysis of Genetic Relationship among Korean Native Orostachys Species Using RAPD (RAPD를 이용한 자생 바위솔속(Orostachys) 식물의 유연관계 분석)

  • Lee, Byoung Ae;Kim, Hag Hyun;Cho, Yong Gu;Lee, Cheol Hee
    • Horticultural Science & Technology
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    • v.19 no.2
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    • pp.159-162
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    • 2001
  • The genetic relationships of Korean native Orostachys species collected from various regions were analysed using random amplified polymorphic DNA (RAPD) method. Eighteen Orostachys species analysed with UPGMA were clustered into three groups A, B, and C. However, four species were not clustered into any group. O. iwarenge species in group A (No. 18-No. 22) showed low similarity with 66.3-73.9% according to the cluster analysis. O. malacophyllus species in group B (No. 12-No. 17) showed low similarity with 66.7-83.7% according to the cluster analysis. The similarity coefficient value of O. japonicus (No. 3-No. 8) except Anmyeondo collected variety (No. 9) showed higher level with 84.2-92.3% than O. iwarenge or O. malacophyllus. Therefore, O. japonicus is thought to be genetically stable, and have less regional variation.

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Analysis of Genetic Polymorphisms and Similarity Using Random Amplified Polymorphic DNAs in Cattle (RAPD기법을 이용한 축우의 유전적 다형성과 유사도 분석)

  • Lee, S.H.;Seo, K.W.;Kwon, I.;Sung, C.K.;Kim, S.K.;Sang, B.C.
    • Korean Journal of Agricultural Science
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    • v.26 no.2
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    • pp.39-48
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    • 1999
  • This study was carried out to investigate the difference and genetic similarity at the level of molecular genetics. Genomic DNA was extracted from blood of Holstein, Korean cattle, Charolais, and hybrid between Korean cattle and charolais and RAPD(random amplified polymorphic DNAs) was analyzed by PCR(polymerase chain reaction). After genetic similarity value from different breeds are analyzed, genetic similarity was estimated by UPGMA(unweighted pair-group method using average). The results obtained from this study can be summarized as follows: 1. When genomic DNA which was extracted from different breeds was subjected to electrophoresis on 1.5% agarose gel, bigger than 12.2kb was appeared. Ratio by absorbance of $A_{260}/A_{280}$ was 1.75~2.10, indicating that genomic DNA was quite pure for RAPD analysis. 2. Different band patterns by RAPD were appeared according to the breeds in cattle. The best primer used to distinguish Holstein from other breeds was 5'-GAC CGC TTG T-3'. 3. A 340bp fragment was amplified in $33.0^{\circ}C$ of annealing temperature for the Holstein and Charolais breeds, but any amplification was not occurred in this annealing temperature for Korean cattle and hybrid. In addition, a 340bp fragment was amplified in $37.5^{\circ}C$ of annealing temperature for the Holstein and Korean cattle, but any amplification was not occurred in this annealing temperature for Charolais and hybrid. For the reaction of PCR. $37.5^{\circ}C$ and $33.0^{\circ}C$ of annealing temperature was shown to be best for genetic marker identification from Holstein, Charolais, and hybrid between Korean cattle and Charolais. 4. When genetic similarity from different breeds are analyzed at the both temperature of $33.0^{\circ}C$ and $37.5^{\circ}C$, the genetic similarity value of Holstein and Korean cattle, Holstein and Charolais, Korean cattle and Charolais, and Korean cattle and hybrid were 0.666~0.777, 0.615~0.666, 0.400~0.461 and 0.857~0.888, respectively. 5. It could be concluded that different breeds are capable of distinguishing by RAPD used random primer 5'-GAC CGC TTG T-3', genetic similarity from different breeds was appeared the higher genetic similarity value of Korean cattle and Charolais than that of Holstein between Korean cattle and Charolais by UPGMA.

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Analyses of Genetic Relationships of Rhizoctonia solani Isolates from Various Crop Species and Rapid Identification of Anastomosis Groups with RAPD Method (각종 작물에서 분리한 R. solani 균주들의 RAPD를 이용한 종내 그룹의 유전적 유연관계 분석 및 AGs 신속 간이동정)

  • Lee, Youn-Su;Choi, Hei-Sun;Kim, Kyoung-Su;Woo, Su-Jin;Kang, Won-Hee;Kim, Myoung-Jo;Shim, Jae-Ouk;Lee, Min-Woong
    • The Korean Journal of Mycology
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    • v.26 no.3 s.86
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    • pp.373-379
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    • 1998
  • Rhizoctonia solani [Thanatephorus cucumeris (frank) Donk], one of the major soil-borne plant pathogens with world-wide distribution, can cause great damages on various crops. In Korea, sheath blight on rice caused by this pathogen is the major concern, and active studies on this pathogen have been performed. However, most of these studies were concerned with pathogenicity of the isolates instead of molecular analyses of different AGs of R. solani. Therefore, in this study, thirty isolates of Rhizoctonia solani collected from various sources were used for the analyses of genetic relationships among themselves and for the rapid anastomosis grouping with RAPD method. As a result, thirty isolates of known and unknown AGs were grouped into five subgroups and each group included AG-1, AG-2, AG-3, AG-4, and AG-5. RS-1 isolate was found to be closely related to AG-5. Isolates RS-4, RS-14, RS-17, and RS-16 were found to be closely related to AG-2-2(III B). Isolate RS-13 was closely related to AG-4, isolates RS-8 and RS-10 were closely related to AG-1(I B), and isolates RS-7 and RS-21 were closely related to AG-2-2(IV). Isolate RS-19 was closely related to AG-1(I C), and isolates RS-3, RS-5, RS-18, RS-6, and RS-15 were found to be closely related to AG-1.

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Studies on Genetic Stability of Micropropagated Plants and, Reintroduction in an Endemic and Endangered Taxon: Syzygium travancoricum Gamble (Myrtacae)

  • Ajith Anand
    • Journal of Plant Biotechnology
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    • v.5 no.4
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    • pp.201-207
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    • 2003
  • Tissue culture techniques arguably are an important approach for ex situ conservation of rare and endangered plant species. However, there is utmost importance on maintaining the genetic integrity of the introduced plants especially in tree species. To examine the genetic integrity of the micropropagated plants, we randomly screened few hardened plants of Syzygium travancoricum, a critically endangered tree taxon, using Randomly Amplified Polymorphic DNA (RAPD) markers. Twenty-three random. primers were tried and twenty-five polymorphic loci were identified. The dendrogram based on the Unweighted Pair-Group Method Arithmetic Average and Nei's similarity index depicted about 97% homology between the mother plants and micropropagated plants. Further, an attempt was made to reintroduce the micropropagated plants in the wild. Over three hundred small trees could be successfully established.

An Efficient Identification of 68 Apple Cultivars Using a Cultivar Identification Diagram (CID) Strategy and RAPD Markers

  • Wang, Wenyan;Wang, Kun;Liu, Fengzhi;Fang, Jinggui
    • Horticultural Science & Technology
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    • v.30 no.5
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    • pp.549-556
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    • 2012
  • The study aimed to establish an efficient tool for cultivar identification and characterization being the first steps of apple introduction and improvement program. We utilized a method to efficiently record DNA molecular fingerprints of plant individuals genotyped by RAPD, which could be used as efficient reference information for quick plant identification. Ten of sixty 11-mer primers were screened to identify the 68 apple genotypes which could be distinguished by a combination of several primers. All cultivars were easily identified by the corresponding primers marked on the cultivar identification diagram (CID). The results indicated that the CID strategy developed and employed in the apple cultivar identification could be vital in the utilization of DNA marker in other plants as well as the development of the apple industry.

Application of RAPD Methods in Meat for Beef Breed Identification

  • Choy, Y.H.;Oh, S.J.;Kang, J.O.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.12
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    • pp.1655-1658
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    • 2001
  • Bovine genome samples were collected from meat of three different beef breeds (Hanwoo, Holstein and imported beef breed) that are commercially merchandized in Korean beef market. Operon B (OPB)-kits were used as random primers (3, 7, 10, 11, 12, 14) in random amplified polymorphic DNA (RAPD) method on whole genome. Each primer provided characteristic bands that were highly polymorphic. Each single primer could provide relatively efficient polymorphic band patterns among breeds. However, use of two or more primers in combination is recommended to improve resolution of experiments with higher molecular weight bands of DNA. In our experiments, OPB-11 resolved well between beef cattle breeds and Holstein. And OPB-7, 12 and 14 could be combined with OPB-11 to identify Hanwoo beef from the other two kinds of beef.

Random Amplified Polymorphic DNA (RAPD) Identification of Genetic Variation in Chlorella species

  • CHO Jung Jong;KIM Yong-Tae;HUR Sung Bum;KIM Young Tae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.6
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    • pp.761-769
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    • 1996
  • The random amplified polymorphic DNA (RAPD) technique was used to characterize 18 reference strains of microalgae, mostly Chlorella species, collected from various localities around Korea peninsular. Eighteen strains consist of four genera of the family marine Chlorella from 12 samples, two genera of fresh water Chlorella from three samples, and three genera on Nannochloris. Twenty 10-mer anonymous primers were screened for amplification of genomic DNA extracted from samples using the CTAB extraction method. Nineteen of these oligonucleotide primers were positive or band producing. Three of 20 random primers (OPA 10, OPA 12, and OPA 18) resulted in both clear band and a high degree of reproducibility and showed some potential to be used to discriminate individual samples of both genetically hetero-and homogeneous populations, in determining phylogenetic relationships between species within a genus and developing individual fingerprints for each samples.

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