• Title/Summary/Keyword: PCR-RAPD marker

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RAPD Pattern of Ginseng(Panax ginseng C.A. Meyer) Lines Containing High Level of Ginsenoside

  • Kang, Tae-Jin;Kim, Se-Young;Rho, Yeong-Deok;Deok-Chun
    • Plant Resources
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    • v.6 no.3
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    • pp.170-174
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    • 2003
  • The important component for medical effect in ginseng is ginsenoside. Korea Ginseng & Tobacco Research Institute contains approximately 200 lines produced by inbred selection. It is assumed that ginseng lines containing high level of ginsenoside should be included in those lines. Besides, new breeding methods such as cell line selection in vitro and hairy root were recently developed. Therefore, this study was carried out to detect genes related to ginsenoside, and to use it for selection marker to select and distribute lines containing high level of ginsenoside. DNA was extracted from both ginseng roots and hairy roots, and the difference between the line containing high ginsenoside(KG101) and normal ginsenoside(KG103) were analysed. As a result, 28 out of 36 primers showed bands, and many primers showed band difference between ginseng lines. It is considered that the bands should be analysed using DNA sequence comparison to check if those are related to ginsenoside. In case of hairy roots of ginseng, almost no differences were found between two lines.

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Genetic Characteristics and Virulence Factors of Pandemic Vibrio parahaemolyticus Isolated in South Korea and Japan (한국과 일본에서 유행하는 장염비브리오의 병원성 인자와 유전자의 특성)

  • Hong, Seuk-Won;Moon, Ji-Young;Lee, Bok-Kwon;Kim, Yung-Bu
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.386-395
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    • 2007
  • A total of 27 strains of Vibrio parahaemolyticus (18 strains isolated from Korea and 9 strains from Japan) were serotyped and examined for biochemical characteristics, antimicrobial susceptibility patterns, cytotoxicity assay, thermostable direct hemolysin (TDH) production and molecular epidemiology. Using polymerase chain reaction (PCR) method and DNA probe hybridization method, the strains were tested for toxR, tdh, trh and ORF 8 genes. The V. parahaemolyticus isolated from patients were belonged to 8 different serotypes : O3:K6, O1:K38, O3:K57, O4:K9, O4:Kl2, O4:K68, O5:Kl5 and O6:K46. Urease-positive strain possessed the trh gene, and conversely, urease-negative strains lacked the gene, indicating that urease production by V. parahemolyticus strains strongly correlates with the possession of the trh gene. Most strains showed multiple resistant to more than three antibiotics and the antibiogram could be classified into 6 group (I to VI). All of the O3:K6 strains isolated in South Korea and Japan producted TDH at high levels. The TDH titers ranged between 256 and 2.048, and the average titer was 1009. To distinguish the new and increasingly common V. parahaemolyticus strains from clinical isolates, ORF 8 is a useful genetic marker. After Southern hybridization, the HindIII restriction fragment patterns of the tdh gene were grouped one type, respectively. One type showed two bands one of which was 4.3kb and the other was 11.5kb in size. Variation between the O3:K6 serotype are minor when compared to the differences seen with the non O3:K6 strains. The migration patterns of Not I -digested of the total DNA of the O3:K6 strains were similar, and only slight variations were observed between the serotypes. By contrast, the O3:K6 strains and non O3:K6 had markedly different profiles. In conclusion, Random amplified polymorphic DNA (RAPD) profile using appropriate primers was an effective epidemiological marker.

Early Identification of Putative Zygotic Seedlings in Citrus Crosses between 'Morita unshiu' (Citrus. unshiu Marc.) and 'Ponkan' (C. reticulata Blanco) Using RAPD and SRAP (RAPD와 SRAP 방법을 이용한 '성전온주'(C. unshiu Marc.)와 '병감'(C. reticulate Blanco) 교잡실생 식별)

  • Yun, Su-Hyun;Moon, Young-Sun;Jin, Seong-Beom;Kang, In-Kyu;Lee, Dong-Hoon
    • Journal of Life Science
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    • v.21 no.4
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    • pp.502-508
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    • 2011
  • The purpose of this study was to evaluate the methods of identifying zygotic seedlings of crosses between 'Morita unshiu' (Citrus. unshiu Marc.) and 'Ponkan' (C. reticulata Blanco). In order to investigate the frequency and position of zygotic seedlings and to determine the efficiency of zygotic seedling identification, random amplified polymorphic DNA (RAPD) and sequence-related amplified polymorphism (SRAP) were performed using UBC (9, 27, 229, 230, and 254) primers and F4/R27, F7/R14, F12/R10, and F44/R62 primer sets, respectively. A total of 37 putative zygotic seedlings out of 55 individuals were selected by RAPD and SRAP. The F7/R14 primer pair showed a screening ability of 45.5% (25/55), whereas the primer UBC27 revealed the highest efficiency of zygotic seedling identification (50.9%, 28/55). When both UBC27 and F7/R14 were properly used for selection of hybridized seedlings of 'Morita unshiu' (C. unshiu Marc.) and 'Ponkan' (C. reticulata Blanco), screening efficiency was increased to 60% (33/55) for putative zygotic seedlings. Thus, it is possible to select putative hybrid zygotic seedlings in an accurate and effective manner by RAPD and SRAP.

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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Screening and Cloning of RAPD Markers from the W Chromosome of Silkworm, Bombyx mori L.

  • Chen, Keping;Zhang, Chunxia;Yao, Qin;Xu, Qinggang;Tang, Xudong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.2
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    • pp.161-167
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    • 2004
  • Silkworms sex determination drew high attention from researchers. Sex chromosomes on the silkworm are of ZW type for females and ZZ type for males. Chromosome W plays an important role in sex determination. Although several molecular linkage maps have been constructed for silkworm, very few markers are discovered on the W chromosome. In order to look for molecular markers and to further locate the Fern gene on chromosome W, we used genomic DNA from both female and male larvae of a silkworm strain named 937 as PCR templates for RAPD amplification with 200 arbitrary 10-mer primers. The amplification results showed three female-specific bands, namely ${OPG-07_496}, {OPC-15_1,660} and {OPE-18_1,279}$. Further verification, however, revealed no band from OPG-07 and OPC-15 in either sex in the strain 798, but OPE-18 provided female-specific band in the strains Suluan7 and C108, and absent in both males and strain 798. This indicates that the bands from ${OPG-07_496} and {OPC-15_1,660}$ are probably female-specific in strain 937, and the band from OPE-18 was probably amplified from a common segment shared by most strains. The genomic DNAs from OPG-07 and OPC-15 were cloned and sequenced. Sequence analysis showed that the DNAs from OPG-07 and OPC-15 have high identities with the retrotransposable elements, and DNA from OPC-15 contains a portion of sequence which probably encodes an eukaryotic translation initiation factor 4E binding protein (eIF4EBP).

S-haplotypes and Genetic Diversity in 'Danji' Radish (Raphanus sativus L. var. hortensis)

  • Ahn, Yulkyun;Kim, Hyukjun;Han, Dongyeop;Park, Younghoon
    • Horticultural Science & Technology
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    • v.32 no.2
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    • pp.210-216
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    • 2014
  • The distribution of S-haplotypes and genetic relationships were evaluated for 47 accessions of 'Danji' radish (Raphanus sativus L. var. hortensis Baker f. gigantissimus Makino) originating from Jeju Island in South Korea. A total of 22 S-haplotype-specific SCAR markers for the S locus glycoprotein (SLG) and S receptor kinase (SRK) loci were tested, and six primer sets amplified locus-specific PCR fragments from at least one 'Danji' radish accession. S5 and S21 alleles atthe SLG locus were the most frequently distributed, and detected from 87.5% and 64.6% of the accessions, respectively. The frequency of the class-II haplotype at the SLG locus was 75%, more frequent than the class-I haplotype. The S23 allele at the SRK locus was detected from 7 accessions. Grouping of the accessions based on S-allele composition revealed three major groups, while 8 accessions showed a unique allelic composition. The genetic diversity of 47 'Danji' radishes and 1 'Gwandong' radish were also evaluated with 38 RAPD primers. A total of 312 bands were scored, and showed that 138 bands (44.2%) were monomorphic among the accessions, whereas 174 (55.8%) bands were polymorphic. Polymorphism rates ranged from 0.2 to 1.0, indicating significant variations in detecting polymorphism across RAPD primers. The genetic similarity coefficients among all pairs of the 48accessions varied from 0.62 to 0.93, and 42% of the comparisons exhibited values higher than 0.85. All the cultivars could be distinguished based on the DNA fingerprints revealed by RAPD. The comparisons between the dendrograms based on S-haplotypes and RAPDs indicate an unrelated and sporadic distribution for several accessions; however, there was a tendency for accessions with the same S-allelic composition to group into the same cluster.

The Relative Identification of C. officinale and L. chuanxiong by PCR-Mediated Fingerprinting (천궁류(川芎類) 한약재의 유전자 감식 연구)

  • Choi, Ho-Young;Kim, Dong-Wook;Kim, Dong-Eun;Suh, Young-Bae;Ham, In-Hye
    • The Korea Journal of Herbology
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    • v.20 no.4
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    • pp.151-161
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    • 2005
  • Objectives : Our research purpose is to establish the standard identification analysis on C. officinale and L. chuanxiong in Korea and China by PCR-mediated fingerprinting. Methods : The Restriction Fragment Length Polymorphism (RFLP) and Randomly Amplified Polymorphic DNA (RAPD) method was used on Internal Transcribed Spacer (ITS) regions and rbcL regions to compare and discriminate genes extracted from crude drugs as C. officinale and L. chuanxiong in Korea and China. Results : L. chuanxiong Korea and China have very similar polymorphism, whereas L. chuanxiong in Korea and C. officinale have very different polymorphism in RFLP. And restriction enzymes AluI and SacI forms the specific fragment band only in C. officinale, they can be used as RFLP marker on ITS regions to discriminate among the species. Conclusions : The results could be applied in discriminating crude drugs among C. officinale and L. chuanxiong in Korea and China. Also they could be used in controlling drug quality, preserving medicinal plants, and improving plant description.

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Geographic Variations and DNA Polymorphisms in Gizzard-shad (Konosirus punctatus) (전어 (Konosirus punctatus)의 지리적 변이와 DNA 다형성)

  • Park, Su-Young;Kim, Jong-Yeon;Yoon, Jong-Man
    • Korean Journal of Ichthyology
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    • v.18 no.4
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    • pp.300-310
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    • 2006
  • Genomic DNA isolated from three geographical gizzard-shad (Konosirus punctatus) populations in Seocheon (SC), Busan (BS) and Gochang (GC) collected in the West Sea and the southern sea, respectively, off the Korean Peninsula, were PCR-amplified repeatedly. Eight selected decamer and 20-mer primers generated a total of 713 loci in the SC population, 791 in the BS population, and 732 in the GC population, with a DNA fragment size ranging from 100 bp to 2,800 bp. We identified 50 unique loci for the SC population, 70 unique loci for the BS population and 130 for the GC population: 120 shared loci for the three populations. There were 108 specific loci (15.1%) for the SC population, 74 (9.4%) for the BS population, and 67 (9.2%) for the GC population. Eight primers also generated 48 polymorphic loci (6.7%) for the SC population, 26 (3.3%) for the BS population, and 16 (2.2%) for the GC population. The similarity matrix ranged from 0.756 to 0.936 for the SC population, from 0.800 to 0.938 for the BS population, and from 0.731 to 0.959 for the GC population. The dendrogram obtained by the eight primers indicates three genetic clusters: cluster 1 (SEOCHEON 01~SEOCHEON 10), cluster 2 (BUSAN 11~BUSAN 20 and GOCHANG 23~GOCHANG 24), and cluster 3 (GOCHANG 21, 22, 25, 26, 27, 28, 29 and 30). As stated above, some individuals of the GC population appear to belong in BS population. When seeing this result, it was thought with the fact that some individuals of 2 populations seem to come and go partially. Thus, RAPD-PCR analysis revealed a significant genetic distance between the three geographical gizzard-shad populations. Using various decamer and 20-mer primers, RAPD-PCR may be applied to identify specific/polymorphic markers that are particular to a species and geographic population, and to define genetic diversity, polymorphisms, and similarities among geographical gizzard-shad populations.

An Overview for Molecular Markers in Plants (식물에서 분자 마커의 동향)

  • Huh, Man Kyu
    • Journal of Life Science
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    • v.25 no.7
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    • pp.839-848
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    • 2015
  • A molecular marker is a molecule contained within a sample taken from an organism or other matter. The development of molecular techniques for genetic analysis has led to a great contribution to our knowledge of plant genetics and our understanding of the structure and behavior of various genomes in plants. Recently, functional molecular markers have been developed to detect the presence of major genes from the analysis of pedigreed data in absence of molecular information. DNA markers have developed into many systems based on different polymorphism-detecting techniques or methods such as RFLP, AFLP, RAPD, SSR, SNP, etc. A new class of very useful DNA markers called genic molecular markers utilizing the ever-increasing archives of gene sequence information being accumulated under the EST sequencing projects on a large number of plant species. Functional markers are derived from polymorphic sequences, and are more likely to be involved in phenotypic trait variation. Based on this conceptual framework, the marker systems discussed below are all (gene)-targeted markers, which have the potential to become functional. These markers being part of the cDNA/EST-sequences, are expected to represent the functional component of the genome i.e., gene(s), in contrast to all other random DNA based markers that are developed/generated from the anonymous genomic DNA sequences/domains irrespective of their genic content/information. Especially I sited Poczai et al’ reviews, advances in plant gene-targeted and functional markers. Their reviews may be some useful information to study molecular markers in plants.

An Introduction to Microsatellite Development and Analysis (Microsatellite 개발 및 분석법에 대한 소개)

  • Yun Young-Eun;Yu Jeong-Nam;Lee Byoung-Yoon;Kwak Myounghai
    • Korean Journal of Plant Taxonomy
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    • v.41 no.4
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    • pp.299-314
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    • 2011
  • The choice of molecular markers is the first step when selecting experimental plans in the field of population genetics. The popular molecular markers in population genetic studies are mainly allozyme, RAPD, RFLP, AFLP, microsatellite, SNP and ISSR. Among these, microsatellites are frequently found in nuclear, chloroplast and mitochondrial genome, showing a high level of polymorphism and nuclear microsatellites are codominant. Thus, it is a favorable molecular marker for population structure analyses and genetic diversity studies. Microsatellites are composed of tandem repeated 1~6 base pair nucleotide motifs and can be easily amplified by PCR reactions using locus specific primers. Because microsatellites have low cross-species transferability, however, they are only applicable between phylogenetically close species. In wild plants, the lack of genomic information and the high development cost of the microsatellite obstruct the wider use of microsatellites in plant population genetics research. In this review, we introduce the basis for microsatellite markers, the development process, and analytical methods as well as evolutionary models and their applications. In addition, possible genotyping errors which lead to erroneous conclusions are discussed.