• Title/Summary/Keyword: random primer

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Serotypes and genotypes of Salmonella isolates from slaughtered pigs (도축돈에서 분리된 살모넬라의 혈청형 및 유전형)

  • Choi, Won-Zong;Jung, Ji-Hun;Won, Ho-Keun;Kang, Zheng-Wu;Hahn, Tae-Wook
    • Korean Journal of Veterinary Service
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    • v.31 no.1
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    • pp.1-16
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    • 2008
  • Salmonella infections cause the disease in pigs but also some zoonotic Salmonella serotypes can be transmitted to human through swine products, resulting in food poisoning. The objective of this study was to investigate the bacteriological prevalence and detection of invA gene using Salmonella specific polymerase chain reaction (PCR), the epidemiological characteristics related to Salmonella strains cultured from pig samples in Gangwon areas using serotyping, random amplified polymorphic DNA (RAPD) and pulsed field gel electrophoresis (PFGE) methods. During the period of November 2001 through April 2002, 1,174 ileocecal lymph node were collected from the slaughtered pigs raised in 38 farms located in Gangwon province. The samples were submerged in boiling water and macerated in saline and lymph node homogenates were inoculated into Tetrathionate broth with iodine (TTB, Difco, 0.5% iodine was added) for enrichment growth. Then additional tests were performed using several mediums, and suspects were identified by API 20E kit (BioMerieux) and PCR. Of total 1,174 samples from 38 farms, 44 (3.7%) were isolated as Salmonella spp from 13 farms (34.2%). Of 44 isolates, 31 were in Yangyang region, followed by 9 in Goseong, 2 in both Gangneung and Sokcho. However, there was no difference in regional isolation frequency. All isolates have a 521bp amplified product in Salmonella specific PCR with primer invA which encodes in proteins for invasion of epithelial cells. Of 44 recovered serotypes, 23 (52.3%) were S Eingedi, 10 (22.7%) S Schwarzengrund, 9 (20.5%) S Typhimurium, and 2 (4.5%) S Mbandaka. In RAPD analysis, there appeared to be unique bands distinguishing each serotype, although similarities exist between the different serotypes. Four serotypes of 44 Salmonella isolates appeared to fall into 14 different RAPD types. In PFGE analysis, 9 S Typhimurium were tested with XbaI enzyme and SpeI enzyme. The combination of results obtained with two enzymes subdivided the 9 S Typhimurium into 4 PFGE types.

Genetic Variation in Among Cultivated Field Populations of Korean Ginseng(Panax ginseng C.A.Meyer) Using RAPD (RAPD marker를 이용한 고려인삼(Panax ginseng C.A.Meyer)의 유전적 변이 분석)

  • 차선경;김영창;최재을;최장선;강권규
    • Korean Journal of Plant Resources
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    • v.16 no.3
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    • pp.251-256
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    • 2003
  • Genetic variation in field grown Korean ginseng(Panax ginseng C.A.Meyer) was evaluated using random amplified polymorphic(RAPD) markers. (This experiment was carried to collect the local native from farm of Chungnam National University in Korea in order to investigate genetic variation.) Some morphological characters showed considerable variation ranging 22 to 68cm in plant hight, 10 to 38mm in root diameter, 16 to 86g in root weight, and culum color and flowering date, respectively. Ten RAPD primers out of the 32 which produced reproducible bands in 662 Korean ginseng plants were selected for the further study. The total number of bands generated by 10 primers were 108 and among them 103 were polymorphic among the 662 plants with the polymorphism ratio of 94.5%. A total of 662 plants were classified into 16 groups based on polymorphic data with an URP 05 primer.

Identification of Arbuscular Mycorrhizal Fungi Colonizing Panax ginseng Using 18S rDNA Sequence (18S rDNA를 이용한 인삼(Panax ginseng)의 내생균근 균의 동정)

  • Eo, Ju-Kyeong;Kim, Dong-Hun;Jeong, Hyeon-Suk;Eom, Ahn-Heum
    • Applied Biological Chemistry
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    • v.47 no.2
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    • pp.182-186
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    • 2004
  • Morphological observation of roots and molecular technique were used to investigate the symbiotic relationships between arbuscular mycorrhizal (AM) fungi and ginseng roots. Korean ginseng, Panax ginseng, was collected from 8 sites in Korea. Colonization pattern of AM fungi in ginseng roots was determined as an Arum type under light microscopes. Nested PCR using AM fungal specific primers was employed to amplify a partial region on 18s rDNA of AM fungi from the root extracted mixed DNA. The amplified DNA was cloned and analyzed by random fragment length polymorphism (RFLP) with restriction enzymes, AluI, HinfI and AsuC21. One from each RFLP pattern was selected for sequencing. A total 16 clones were sequenced and identified as 2 species of AM fungi; Paraglomus brasilianum and Glomus spurcum. Paramglomus brasilianum was found from most of the ginseng roots, in this syudy suggesting that this species of AM fungi could have specific relationship with the ginseng root. Possible roles of AM fungal species in ginseng roots are discussed.

Identification of a New Potyvirus, Keunjorong mosaic virus in Cynanchum wilfordii and C. auriculatum (큰조롱과 넓은잎 큰조롱에서 신종 포티바이러스(큰조롱모자이크바이러스)의 동정)

  • Lee, Joo-Hee;Park, Seok-Jin;Nam, Moon;Kim, Min-Ja;Lee, Jae-Bong;Sohn, Hyoung-Rac;Choi, Hong-Soo;Kim, Jeong-Soo;Lee, Jun-Seong;Moon, Jae-Sun;Lee, Su-Heon
    • Research in Plant Disease
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    • v.16 no.3
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    • pp.238-246
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    • 2010
  • In 2006 fall, a preliminary survey of viruses in two important medicinal plants, Cynanchum wilfordii and C. auriculatum, was conducted on the experimental fields at the Agricultural Research and Extension Services of Chungbuk province in Korea. On each experimental fields, percentage of virus infection was ranged from 20 to 80%, and especially an average of disease incidence propagated by roots was twice higher than that by seeds. The various symptoms were observed in Cynanchum spp. plants, such as mosaic, mottle, necrosis, yellowing, chlorotic spot and malformation etc. In electron microscopic examination of crude sap extracts, filamentous rod particles with 390-730 nm were observed in most samples. The virus particles were purified from the leaves of C. wilfordii with typical mosaic symptom, and the viral RNA was extracted from this sample containing 430-845 nm long filamentous rod. To identify the viruses, reverse transcription followed by PCR with random primers was carried out. The putative sequences of P3 and coat protein of potyvirus were obtained. From a BLAST of the two sequences, they showed 26-38% and 62-72% identities to potyviruses, respectively. In SDS-PAGE analysis, the subunit of coat protein was approximately 30.3 kDa, close to the coat protein of potyvirus. In bioassay with 21 species in 7 families, Chenopodium quinoa showed local lesion on inoculated leave and chlorotic spot on upper leave, but the others were not infected. RT-PCR detection using specific primer of C. wilfordii and C. auriculatum samples, all of 24 samples with virus symptom was positive, and five out of seven samples without virus symptom were also positive. On the basis of these data, the virus could be considered as a new member of potyvirus. We suggested that the name of the virus was Keunjorong mosaic virus (KjMV) after the common Korean name of C. wilfordii.

Characteristics and breeding of a new cultivar Pleurotus eryngii, Song-A (큰느타리버섯 신품종 '송아'의 육성 및 그 특성)

  • Shin, Pyung-Gyun;Park, Yun-Jung;Yoo, Young-Bok;Kong, Won-Sik;Jang, Kab-Yeul;Cheong, Jong-Cheon;Oh, Se-Jong;Lee, Keum-Hee
    • Journal of Mushroom
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    • v.9 no.2
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    • pp.59-62
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    • 2011
  • To develop a new cultivar of King oyster mushroom(Pleurotus eryngii), G09-21 as parental strain was selected by the method of Di-mon crossing between monokaryotic strains derived from ASI 2824(Keunneutari No.2) and dikaryotic strain ASI 2887(Aeryni 3). The Pe21-51($G09-21-10{\times}2844-11$) was shown the best cultural characteristics, selected to be a new cultivar and named as 'Song-A'. The 'Song-A' was formed incompatibility line distinctly in the confrontation growth of parental strains Keunneutari No.2, Aeryni 3 and ASI 2844. The optimum temperature for mycelial growth, fruiting body development and pH arrange were $25{\sim}30^{\circ}C$, $14{\sim}16^{\circ}C$ and pH5~8, respectively. Fruiting body production per bottle was about $94.7{\pm}29.5$ g which is almost 106% quantity compared to that of other cultuvar Keunneutari No.2. And also the stip is thick and long but the number of available stipe is few. Analysis of the genetic characteristics of the new cultivar 'Song-A' showed a different DNA profile as that of the control strains, Keunneutari No.2, Aeryni 3 and ASI 2844, when RAPD(Random Amplified Polymorphic DNA) primers URP4 and 7 were used. This new cultivar 'Song-A' of Pleurotus eryngii is characterized by a small number of primordia formation and the stip is thick and long. Therefore, we expect that this new strain will save of labor and cost by without culling work.

Characteristics and breeding of a new multi-generation oyster mushroom (Pleurotus ostreatus) variety 'Dagul' (다발성 신품종 느타리 '다굴'의 육성 및 자실체 특성)

  • Shin, Pyung-Gyun;Kim, Hee-Jung;Choi, Chan-Sik;Yoo, Young-Bok;Kong, Won-Sik;Jang, Kab-Yeul;Oh, Youn-Lee;Cheong, Jong-Chun;Suh, Jang-Sun;Oh, Se Jong;Lee, Keum-Hee
    • Journal of Mushroom
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    • v.11 no.3
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    • pp.154-158
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    • 2013
  • To develop a new variety of oyster mushroom(Pleurotus ostreatus), parental strains was selected by the method of Mon-Mon crossing between monokaryotic strains derived from ASI 2596(Suhan No.3) and ASI 2782(Black pileus mutant). The SB-73(ASI 2596-11 x 2782-8) was shown the best cultural characteristics, selected to be a new variety and named as 'Dagul'. The 'Dagul' was formed incompatibility line distinctly in the confrontation growth of parental strains Suhan No.3 and ASI 2782. The optimum temperature for mycelial growth, fruiting body development and pH arrange were $25{\sim}30^{\circ}C$, $14{\sim}17^{\circ}C$ and pH5~8, respectively. Fruiting body production per bottle was about $68.0{\pm}24.1$ g which is almost 115% quantity compared to that of other variety Suhan No.3. And also the stipe is long and individual generation is multiple. Analysis of the genetic characteristics of the new variety 'Dagul' showed different DNA bands as that of the control strains, Suhan No.3 and ASI 2782, when RAPD(Random Amplified Polymorphic DNA) primers URP7 and Rcb1 were used. This new variety 'Dagul' of oyster mushroom is characterized by multiple of individual generation and the stipe is long. We therefore expect that this new strain will increase of the income by cultivation of field.

MORPHOLOGIC CHANGE OF DENTIN SURFACE ACCORDING TO THE DIFFERENCE IN CONCENTRATION AND APPLICATION TIME OF PHOSPHORIC ACID (인산용액의 농도 및 적용시간 차이에 따른 상아질 표면의 형태적 변화)

  • Kim, Myeong-Su;Ohn, Young-Seok;Lee, Kwang-Won;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.23 no.1
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    • pp.141-161
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    • 1998
  • The depth and patterns of demineralization according to the difference in concentration and application time of phosphoric acid were observed through the transmission electron microscope, and shear bond strengths to the acid -conditioned dentin were then measured and compared with the TEM results. To investigate the influence of polymer addition into the phosphoric acid and the effect of difference in concentration and application time of the acid, the specimens were randomly divided into 9 groups. Among the specimens, the exposed dentin surfaces were acid-conditioned with 10% polymer-thickened phosphoric acid(All Bond 2, Bisco, U.S.A.) and aqueous 10%, 20%, 30%, 40% phosphoric acid for 20 seconds, The rest of the specimens were acid-conditioned with 10% phosphoric acid for 15s, 30s, 60s, 120s respectively. The specimens were immersed in 4% glutaraldehyde in 0.1M sodium cacodylate buffer and postfixed with 1 % osmium tetroxide without decalcification and then observed under a JEOL Transmission Electron Microscope(JEM 1200 EX II, Japan). After the specimens were acid-conditioned as the above, primer and adhesive resin were applied to blot-dried dentin and shear bond strengths were then measured and analysed. The results were as follows : 1. The intertubular demineralization depth of 4.0-$5.0{\mu}m$ in 10% polymer-thickened phosphoric acid gels was similar or slightly deeper than that of 4.0-$4.5{\mu}m$ in aqueous 10% phosphoric acid solution. 2. The intertubular demineralization depth of aqueous 20%, 30% and 40% phosphoric acid solution was 6.5-$7.0{\mu}m$, 6.5-$7.5{\mu}m$ and 9.0-$15.0{\mu}m$ respectively. It showed that the depth of dentin demineralization is partly related to the concentration of phosphoric acid solution. 3. The intertubular demineralization depth of aqueous 10% phosphoric acid solution in application time for 15s, 30s, 60s and 120s was 2.5-$3.0{\mu}m$, 4.0-$6.0{\mu}m$, 6.5-$7.0{\mu}m$ and 8.5-$14.0{\mu}m$ respectively. It showed that the depth of dentin demineralization is directly related to the application time of phosphoric acid solution. 4. The partially demineralized dentin layer between demineralized collagen layer and unaffected dentin was showed to a width of 0.5-$1.0{\mu}m$ in lower concentration groups treated with aqueous 10% phosphoric acid for 20s, 60s, 120s and 20% phosphoric acid for 20s. 5. The demineralization effect at the border of intertubular-peritubular junction was less evident than that in the peritubular and intertubular dentin. The collagen fibers in the intertubular dentin had a random orientation, whereas those that lined the tubules were circumferentially aligned. The cross-linkage of dentinal collagen in demineralized collagen layer was clearly seen. 6. A statistically significant difference of bond strengths according to the difference in phosphoric acid concentration did not exist among the groups treated with 10%, 20%, 30% and 40% acid solution (P>0.05). However, bond strengths to the treated dentin with 10% phosphoric acid solution for 30s were significantly higher than that for 120s (P<0.05).

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Analysis of Genetic Diversity of Apple Cultivars Using RAPD and SSR Markers (RAPD와 SSR 마커를 이용한 사과 품종의 유전적 다양성 분석)

  • Cho, Kang-Hee;Heo, Seong;Kim, Jeong-Hee;Shin, Il Sheob;Han, Sang Eun;Kim, Se Hee;Kim, Dae-Hyun;Kim, Hyun Ran
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.525-533
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    • 2010
  • In this study, random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) analyses were utilized for evaluation of genetic diversity of 34 Korean bred and introduced apple cultivars. Thirty-seven RAPD primers detected a total of 193 polymorphic bands (36.2%) with an average of 5.6. Twenty-six SSR markers generated a total of 112 alleles with an average 4.3 alleles per locus. Genetic diversity of 34 cultivars estimated by polymorphic information content (PIC) value ranged from 0.536 (CH03d12) to 0.952 (CH04c06) with an average of 0.843. By UPGMA (unweighted pair-group method arithmetic average) cluster analysis with 305 polymorphic bands, the apple cultivars were classified four groups by similarity index of 0.640. The 'Seokwang' was included in group I. Group II consisted of 12 cultivars which have 'Golden Delicious' in their pedigree, with the exception of 'Spur Earliblaze' and 'Jonathan'. Group III included 13 cultivars which have usually 'Fuji' in their ancestry and bud sport of 'Fuji' cultivars. Group IV consisted of 8 cultivars with 'Hongro', 'Gamhong', and 'Saenara'. Similarity values among the tested apple cultivars ranged from 0.529 to 0.987, and the average similarity value was 0.647. The similarity index was the highest (0.987) between 'Hwarang' and 'Danhong', and the lowest (0.529) between 'Seokwang' and 'Hwarang'. The genetic relationships among the 34 studied apple cultivars were basically consistent with the known pedigree.