• Title/Summary/Keyword: random amplification of polymorphic DNA

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DNA Profiles Analysed by Polymerase Chain Reaction-Random Amplified Polymorphic DNAs in Shortnecked Clam (Ruditapes philippinarum) Populations

  • Yoon, Jong-Man;Kim, Yong-Ho;Kim, Jong-Yeon
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2002.08a
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    • pp.172-174
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    • 2002
  • Out of 20 primers, 6 generated a total of 1,11 major and minor RAPD bands, producing approximately 4.2 average polymorphic bands pe primer in shortnecked clam (Ruditapes philippinarum) population from Anmyeondo. The bandsharing value altered form 0.15 to 0.74, with the average of 0.5, as calculated by bandsharing analysis. The RAPD profiles obtained with DNAs of two populations from Anmyeondo and Seocheon, respectively, were considerably different (0.20 and 0.51, respectively). The varying degrees of difference among populations may also be of relevance to the retricted hybridization of wild bivalve. Besides gene mapping and breeding applications, PCR-RAPD system could be very useful for the rapid certification and quality control of seed production and for every projects based on PCR amplification of specific bivalve DNA fragments.

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Genetic Relationahips of the Two Morphorogical Types of Myzus persicae(Homoptera:Aphididae) Collected from Tobacco Plants Based on Random Amplified Polymorphic DNA(RAPD) (연초에서 발생하는 복숭아혹진딧물(Myzus persicae)형태형 2종의 Random Amplified Polymorphic DNA(RAPD)을 이용한 유전적 유연관계 분석)

  • 채순용;이기원;김상석;장영덕
    • Korean journal of applied entomology
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    • v.37 no.1
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    • pp.31-37
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    • 1998
  • Random amplified polymorphic DNA (RAPD) markers were used to analyze genetic similarity among 8 clones of apierous green peach aphid, two types (M. persicae Sulzer and M. nicotianae lack man) classified by their mo~hologi~cahla raters and host preference (Blackman, 1987), collected from tobacco plants. The genetic variation among these clones was evaluated by polymerase chain reaction amplification with 20 random primers. The higher GC contents of primers, the better in amplification efficiency of PCR reaction in general. The genetic similarities among eight aphid clones were analyzed from UPGMA (unweighted pair group average method) cluster analysis based on simple matching coefficient. The range of genetic similarity coefficients was 0.414 to 0.808. The most close relationship among the clones was similarity coefficient of 0.808 between the PG2 and the PG3 clone. The eight aphid clones analyzed were clustered into three groups by the genetic similarity coefficient. The first group, PG1, PG2, PG3 clone including in M. persicae type by their morphological characters and RED clone in M. nicotianae type was clustered at the genetic similarity coefficient of 0.643. The second group, GR1, GR2, BRN in M. nicotianae type was at the 0.636;and the third group was DBR clone in M. persicae type. The results did not indicate any correlation between m&-phological types (M. persicae and M. nicotianae) and RAPD polymorphism. We could not detect any obvious genetic relationships of the two morphological types of the green peach aphid collected from tobacco plants.

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Random amplification of polymorphic DNA typing of Listeria monocytogenes isolates from animal products (축산물유래 Listeria monocytogenes의 RAPD typing)

  • Lee, Chul-Hyun;Son, Won-Geun
    • Korean Journal of Veterinary Research
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    • v.49 no.4
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    • pp.319-328
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    • 2009
  • This study investigated the epidemiology of Listeria (L.) monocytogenes, a food-borne pathogen. The epidemiology of food-borne pathogens is of great importance for clarifying bacterial origin and preventing bacterial contamination and infection. This work examined 68 L. monocytogenes strains, including 11 reference strains and 57 isolates from imported US beef, domestic meats (beef, pork, chicken meat), raw milk, and milk plants. The random amplified polymorphic DNA (RAPD) techniques were optimized to develop a standard molecular epidemiological analysis of L. monocytogenes. There was great genetic variability among the isolates, which produced 24 and 34 RAPD patterns with primer HLWL85 and HLWL74, respectively. The discriminatory power of the RAPD methods with HLWL85 and HLWL74 primer were very high (DI = 0.957; S ${\geq}$ 80%, S ${\geq}$ 95%). Some RAPD types were specific to origin. A few RAPD types were specific for L. monocytogenes strains belonging to a particular serotype. Using the HLWL85 primer, the strains isolated from milk plants could be distinguished from the other strains. And using the HLWL74 primer, the strains isolated from imported beef (US) could be distinguished completely from the other strains.

Limits of Direct PCR Amplification from Seaweeds Using Arbitrary and ITS Primers (해조류로부터 Arbitrary 및 ITS Primer들을 사용한 직접 PCR 유전자 증폭반응의 한계)

  • 김용국;진형주;박선미;진덕희;홍용기
    • Journal of Life Science
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    • v.9 no.1
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    • pp.15-21
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    • 1999
  • The random amplified ploymorphic DNAs (RAPD) assay is a simple and useful tool in identification of appropriate genetic markers, that requires no knowledge of target DNA sequence. RAPD products were generated directly from seaweed tissues, without prior nucleic acid extraction, of Porphyra yezoensis, Ulva pertusa and Undaria pinnatifida. The nuclear rDNA internal transcribed spacer (ITS) fragment however was not amplified directly from the seaweed tissues. Using DNA extracted by the LiCl method, both the ITS and RAPD's have been amplified by the polymerase chain reaction. RAPD of P yezoensis, thallus (n) and conchocelis (2n) produced lots of different polymorphic bands (36-50$\%$) depending on the arbitrary primers used. Difference was also observed between direct tissues amplification and DNA extracts amplification (53-57$\%$). Thus it is important to use the same ploidy of tissue for DNA extraction and as a RAPD template.

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Rapid molecular authentication of three medicinal plant species, Cynanchum wilfordii, Cynanchum auriculatum, and Polygonum multiflorum (Fallopia multiflorum), by the development of RAPD-derived SCAR markers and multiplex-PCR

  • Moon, Byeong-Cheol;Choo, Byung-Kil;Cheon, Myeong-Sook;Yoon, Tae-Sook;Ji, Yun-Ui;Kim, Bo-Bae;Lee, A-Young;Kim, Ho-Kyoung
    • Plant Biotechnology Reports
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    • v.4 no.1
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    • pp.1-7
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    • 2010
  • Definitive identification of original plant species is important for standardizing herbal medicine. The herbal medicines Cynanchi Wilfordii Radix (Baekshuoh in Korean and Beishuwu in Chinese) and Polygoni Multiflori Radix (Hashuoh in Korean and Heshuwu in Chinese) are often misidentified in the Korean herbal market due to morphological similarities and similar names. Therefore, we developed a reliable molecular marker for the identification of Cynanchi Wilfordii Radix and Polygoni Multiflori Radix. We used random amplified polymorphic DNA (RAPD) analysis of three plant species, Polygoni multiflorum, Cynanchum wilfordii, and Cynanchum auriculatum, to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed six sequence characterized amplification region (SCAR) markers for distinguishing Polygoni Multiflori Radix and Cynanchi Wilfordii Radix. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR. These SCAR markers can be used for efficient and rapid authentication of these closely related species, and will be useful for preventing the distribution of adulterants.

Assessment of genetic diversity of Typha angustifolia in the development of cattail stands

  • Min, So-Jung;Kim, Heung-Tae;Kim, Jae-Geun
    • Journal of Ecology and Environment
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    • v.35 no.1
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    • pp.27-34
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    • 2012
  • Typha angustifolia has ecological characteristics of clonal growth similar to Phragmites australis. The plant spreads byclonal growth and seed dispersal. In this study, for the three stands which have different settlement age at the Baksilji wetland in Korea, genetic diversity was estimated by random amplification of polymorphic DNA analysis to evaluate the change in genetic diversity of T. angustifolia during stand development in the same population. Stand (ST) 1 was the oldest and ST 4 was the youngest. ST 5 was in a small ditch out of the Baksilji. Although the ST 1, ST 2, and ST 3 did not differ significantly in vegetational or physical environment, the genetic diversity estimated according to Nei's gene diversity (h) and the Shannon index (i) increased in the order of ST 1 < ST 2 < ST 3 contrary to formative age. The genetic diversity of ST 4 was much higher than that of the other three stands. ST 4 has similar abiotic environmental conditions with slight T. angustifolia dominance, and seems to be in the early establishment stage. ST 5 differed from the other stands in vegetational and soil environments, which can result in stressful cattail conditions. Even though the ST 5 stand was not younger than the ST 4 stand, ST 5 showed the highest genetic diversity. Our results indicate that after early settlement of the T. angustifolia population, genetic diversity within the species decreased over time and that the decreasing pattern of genetic diversity within T. angustifolia stands is not likely to occur under stressful conditions.

Construction of a linkage Map in Capsicum annuum L. Using RAPD Markers and Identification of Two QTLs.

  • Yang, Tae-Jin;Kim, Yong-Jae;Park, Hyo-Guen
    • Journal of Plant Biotechnology
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    • v.1 no.2
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    • pp.109-115
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    • 1999
  • A linkage map of Capsicum annuum L. was constructed by random amplified polymorphic DNA (RAPD) markers followed in a backcross population of an intraspecific cross between cultivars HDA210 and Yatsufusa. A total of 420 random primers were tested and 311 polymorphic bands were generated by 158 random primers. Among them, 86 Yatsufusa specific bands generated by 52 primers were examined for mapping. Most bands except three segregated in Mendelian fashion fitting the expected 1:1 ratio. The total length of the map was 533 cM distributed in 15 linkage groups. The map distance between adjacent markers ranged 0 to 32.8 cM, with an average distance of 9.1 cM (63 markers). Some markers were clustered and this may be due to the amplification of a repetitive sequence by the RAPDs. Primer pairs for a sequence characterized amplified region (SCAR) were developed and the segregation scores by the SCAR primers were in accordance with the RAPD data. Two QTL markers for number of axillary shoots and for early flowering were developed. One QTL for early flowering located in the linkage group 3 and explained 61 "io of the phenotypic variation. The other QTL for the number of axillary shoots located in the linkage group 4 explained 55 % of the phenotypic variation.tion.

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Characteristics and breeding of a thermotolerant ear mushroom, Auricularia auricula-judae 'Hyeonyu' (고온적응성 목이버섯 「현유」 육성 및 특성)

  • Lee, Gl-Kwon;Yu, Young-Jin
    • Journal of Mushroom
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    • v.15 no.2
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    • pp.84-87
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    • 2017
  • Although ear mushroom (Auricularia auricula-judae) is cultivated worldwide, there are a limited number of commercial cultivars in Korea. Recent increase in the import of ear mushroom from China threatens the domestic farming. The present study introduces a new thermotolerant cultivar of ear mushroom, designated 'Hyeonyu', developed by mating monokaryons obtained from JBAA11 and CAA1 strains. The optimal growth temperature for Hyeonyu was 26-36?. The periods of primordia formation and fruit-body growth were 27 and 15 days, respectively. The average mushroom yield per bag (1 kg) was 350 g. Random amplification of polymorphic DNA (RAPD) analysis with OPA1 and OPA7 primers identified polymorphic DNA bands between the control, the new variety Hyeonyu, and a Chinese variety.

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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Monitoring of Gentic Variability in Dicofol-susceptible, Dicofol-resistant, and its Reverse-selected Strains of Tetranychus urticae by RAPD-PCR

  • Song, Cheol;Park, Jin-Hee;Kim, Gil-Hah;Kwon, O-Yu;Cho, Kwang-Yun
    • Journal of Life Science
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    • v.9 no.1
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    • pp.14-16
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    • 1999
  • Genetic variability was monitored by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) in dicofol-susceptible (S), dicofol-resistant (R) and its reverse-selected (RS) strains of two-spotted spider mite, of Tetranychus urticae. Before the reverse-selection, RS strain, selected reversely from R strain, was 23-fold resistance ratio at {TEX}$LC_{50}${/TEX} to S strain. The resistance was started to in incline slowly to the resistance level of S strain after one year, and the resistance ratio was 4-fold in the 7 years after then. PCR-amplification of T. urticae DNA showed polymorphism in the amplifications with 12 primers in 100 kinds of arbitrary DNA sequences. RAPD amplification with primer OPR-12 (5`-ACAGGTGCGT-3`) showed amplified bands at 1,000 base pair in the S-and RS-strain, and at 350 base pair in R-strain. The results of polymorphism are genetic variabilities derived from development and selection of resistance in each strain. The peculiarly amplified fragments were guessed to participate in dicofol resistance. From the analysis of genetic similarity, it is inferred the gene composition of S-and RS-strain is much closer than that of R-strain.

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