• Title/Summary/Keyword: randomly amplified polymorphic DNA (RAPD)

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RAPD Analysis on the Species of Pinelliae Tuber (RAPD 방법을 이용한 반하류 한약재의 감별 연구)

  • 배명효;김규열;정유헌;최호영
    • The Journal of Korean Medicine
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    • v.20 no.4
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    • pp.16-22
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    • 2000
  • This study intends to report the significance of several experimental results obtained from analysing the genes extracted from the plants and herbal medicine such as P. temalta (Thunb.) Breit, A. amurense var serratum Nakai, A. erubescens (Wall.) Schott, Pinelliae Tuber and Arisaematis Tuber, mainly by the method of RAPD(randomly amplified polymorphic DNA) and the method of RFLP(restriction fragment length polymorphism) on ITS(internal transcribed spacer) region. Genomic DNA could be extracted from both original plants and dried materials. DNA fragments of P. temata kind and A. amurense kind showed the same aspect separately within the same species under the method of RAPD using random primer, while various aspects(polymorphism) were discovered among different species. In RAPD analysis by uniprimer, common bands were extracted from all types of P. temata in the case of uniprimer #4, which were distinguished from the kind of A. amurense. Other polymorphic bands appeared in between different A. amurense species as well. In the case of uniprimer #11, particular band came out in the kind of P. temata. On the other hand, in the case of uniprimer #5, #6, and #8, various bands(polymorhism) were revealed in both kinds of P. temata and A. amurense. Although further study is needed to ascertain whether these results are due to the differences of species, kinds, or growing place, the results could be used as a scientific method of identifying the substitutes for A. amurense genus. The author believes that as if P. temata class of plants used in this experiment are different among themselves in terms of the shape, size and property, those are clearly a class of P. temata or belong to the same genus.

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Toxin Gene Typing, DNA Fingerprinting, and Antibiogram of Clostridium perfringens Isolated from Livestock Products

  • Lee, Seung-Bae;Choi, Suk-Ho
    • Food Science of Animal Resources
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    • v.26 no.3
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    • pp.394-401
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    • 2006
  • Forty Clostridium perfringens isolates were obtained from twelve animal products, following the examination of eighty six beef, pork, broiler chicken and salami meat products, and eleven milk powder products. There were 21 isolates from salami stored at $25^{\circ}C$, 3 isolates from pork, 4 isolates from beef, 9 isolates from broiler chicken, and 3 isolates from milk powder. Only the cpa gene encoding a toxin among the 5 toxin genes tested (cpa, cpb, etx, iap, and cpe) was detected in all forty isolates, suggesting contamination with C. perfringens type A. DNA fingerprinting analysis using PCR of the tRNA intergenic spacer (tDNA-PCR) and the 16S-23S internal transcribed spacer (ITS-PCR), and randomly amplified polymorphic DNA (RAPD) analysis were attempted to differentiate the isolates. RAPD analysis was the most discriminating method among the three PCR analyses. Isolates from the same products tended to show similar RAPD patterns. Antimicrobial susceptibility tests showed that some isolates from broiler chickens had the same antibiogram with multiple resistance to streptomycin, colistin, and ciprofloxacin. Antibiograms were similar between isolates from the same livestock products, but differed considerably between the products.

Identification of Korean Mountain Cultivated Ginseng by RAPD (RAPD(Random Amplified Polymorphic DNA)를 이용한 장뇌삼의 지역별 품종 구분)

  • Choi, Ji-Young;Lee, Ju-Hee;Lee, Su-Gwang;Kang, Ho-Duck
    • Journal of agriculture & life science
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    • v.43 no.6
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    • pp.35-43
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    • 2009
  • This study was conducted to examine the genetic variations and intraspecific relationships between 9 individuals of Panax ginseng C.A Meyer by using RAPD (Randomly Amplified Polymorphic DNA) analysis. The 34 primers out of 40 random primers were amplified for all tested plants. The 48 (40%) among 244 bands derived from 34 primers shown polymorphism, and the 72 (64%) rest of bands showed similar forms. By regional groups Sangju and Andong samples located in Kyungsang buk-do showed a high similarity. However, Punggi located in Kyungsang buk-do showed higher similarity with Jinan's of Junla buk-do. In this way, it did not show that Panax ginseng from the same area has similarities. In future study we need to more specific molecular phylogenetic analysis such as AFLP technology and gene sequencing with nuclear chloroplast DNA in all samples.

Genetic Relationships Analysis of Asparagus cochinchinensis $L_{OUR}$ Collections by Random Amplified Polymorphic DNA (RAPD에 의한 천문동 수집종의 유연관계 분석)

  • Kang, Chan-Ho;Park, Chun-Bong;Choi, Joung-Sik;Choi, Yeong-Geun
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.5
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    • pp.384-391
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    • 2002
  • To analyze the genetic relationships among 23 accessions of Asparagus cochinensis $L_{OUR}$, random amplified polymorphic DNA(RAPD) analysis was performed using artificially synthesized 10 primers. The range of polymorphism was $42.9{\sim}91.7%$ with an average of 72.9% in 85 randomly and specifically amplified DNA fragments. On the. basis of similarity coefficient analysis by unweighed pairgroup method, arithmetic average method(UPGMA), 23 accessions of Asparagus cochinensis $L_{OUR}$ could be classified into 6 groups at the similarity coefficient value of 0.82. Group I contained 5 accessions, Group II contained 6 accessions, Group III contained 6 accessions, GroupIV contained 2 accessions, Group V contained 2 accessions and Group VI contained 2 accessions. The range of total genetic similarity coefficient value of 23 accessions of Asparagus cochinchinensis $L_{OUR}$ was $0.47{\sim}0.92$ and average value was 0.76. To obtain more exact data from PCR, we also tried to develope enhanced RAPD techniques using Bovine Lacto Transfer Technique Optimizer(BLOTTO). In RAPD analysis of Asparagus cochinensis $L_{OUR}$, we could obtain better RAPD results by adding BLOTTO at a final concentration of 1%.

Species Identification of the Tropical Abalone (Haliotis asinina, Haliotis ovina, and Haliotis varia) in Thailand Using RAPD and SCAR Markers

  • Klinbunga, Sirawut;Amparyup, Piti;Leelatanawit, Rungnapa;Tassanakajon, Anchalee;Hirono, Ikuo;Aoki, Takashi;Jarayabhand, Padermsak;Menasveta, Piamsak
    • BMB Reports
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    • v.37 no.2
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    • pp.213-222
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    • 2004
  • A randomly amplified polymorphic DNA (RAPD) analysis was used to identify the species- and population-specific markers of abalone; Haliotis asinina, H. ovina, and H. varia in Thai waters. Fifteen species-specific and six population-specific RAPD markers were identified. In addition, an 1650 bp band (UBC195) that was restricted to H. ovina from the Gulf of Thailand (east) was also found. All of the specific RAPD markers were cloned and sequenced. Twenty pairs of primers were designed and specificity-tested (N = 12 and 4 for target and non-target species, respectively). Seven primer pairs (CUHA1, 2, 4, 11, 12, 13, and 14) were specifically amplified by H. asinina DNA, whereas a single pair of primers showed specificity with H. ovina (CUHO3) and H. varia (CUHV1), respectively. Four primer pairs, including CUHA2, CUHA12, CUHO3, and CUHV1, were further examined against 216 individuals of abalone (N = 111, 73, and 32, respectively). Results indicated the species-specific nature of all of them, except CUHO3, with the sensitivity of detection of 100 pg and 20 pg of the target DNA template for CUHA2 and CUHA12 and CUHV1, respectively. The species-origin of the frozen, ethanol-preserved, dried, and boiled H. asinina specimens could also be successfully identified by CUHA2.

Optimum Condition of Polymerase Chain Reaction Techniques for Randomly Amplified Polymorphic DNA of Strawberry (딸기의 RAPD를 위한 PCR의 최적조건)

  • 양덕춘;최성민;강태진;이미애;송남현;민병훈
    • Korean Journal of Plant Resources
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    • v.14 no.1
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    • pp.65-70
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    • 2001
  • This study was performed to select marker which can identify genetic variation between mother plant and in vitro cultured plantlets of strawberry by PCR using random primer. When 'Yeobong' DNA extracted was treated with proteinase-K and RNase-H, clear DNA bands were shown. The optimal condition for RAPD in strawberry was to use 50ng of template DNA, 10pmol of primer,37oC of annealing temperature, and 45 cycles of PCR. After establishing above PCR optimal condition, RAPD pattern was investigated by using UBC primers. PCR was performed, and 46 of 90 primers produced PCR product showing 158 total bands. GC content was compared between the primers forming bands and no bands. The GC content showing bands was average 67.4%, whereas primers showing no bands 58%.

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Molecular discrimination of Panax ginseng species

  • Um, Jae-Young;Chung, Hwan-Suck;Kim, Hyun-Ju;Kim, Dae-Ki;Shim, Kyung-Shik;Lee, Kang-Yong;Kim, Jeong-Sook;Choi, Tae-Jin;Kim, Nam-Song;An, Nyeon-Hyoung;Lee, Kang-Min;Lee, Young-Mi;Kim, Jeong-Joong
    • Advances in Traditional Medicine
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    • v.1 no.2
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    • pp.52-58
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    • 2000
  • In order to develop convenient and reproducible methods for identification of ginseng drugs at a DNA level, RAPD (randomly amplified polymorphic DNA) and PCR-RFLP (PCR-Restriction fragment length polymorphism) analysis were applied within Panax species. To authenticate Panax ginseng betvyeen Chinese and Korean ginseng population, RAPD analysis were carried out using 20 mer-random primer. The similarity coefficients among the DNA of ginseng plants analyzed were low, ranging from 0.197 to 0.491. In addition, using PCR-RFLP analysis, very different fingerprints were obtained within Korean ginseng plants. These results suggest that these methods are able to authenticate the concerned Panax species. Broader application of this approach to authenticate other morphologically similar medicinal materials is rationalized.

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Development of Suhan Strain-specific SCAR Marker in Pleurotus ostreatus (느타리 버섯에서 수한 품종 특이 SCAR marker 개발)

  • Seo, Kyoung-In;Jang, Kab-Yeul;Yoo, Young-Bok;Park, Soon-Young;Kim, Kwang-Ho;Kong, Won-Sik
    • The Korean Journal of Mycology
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    • v.39 no.1
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    • pp.31-38
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    • 2011
  • In this study, 81 commercial strains of Pleurotus species cultivated in South Korea were analyzed with randomly amplified polymorphic DNA (RAPD) technique. Sequence characterized amplified region (SCAR) markers were developed by designing from one RAPD polymorhic band specific to Suhan strain. The SCAR primer pair 'S-OPA13-1' amplified a 590-bp fragment in the varieties originated from Suhan strain. The Blast search of S-OPA13-1 showed high homology to the POMFBO1 P. ostreatus cDNA clone MFB02-A05 and Laccaria bicolor S238N-H82. The results showed that this SCAR marker can clearly distinguish Suhan strains from Pleurotus spp.

Development of a Female-associated SCAR Marker in Schisandra nigra Max. (Schisandra nigra Max.에서 암그루에 연관된 SCAR 마커의 개발)

  • Han, Hyo Shim;Jung, Jae Sung
    • Journal of Life Science
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    • v.31 no.6
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    • pp.537-542
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    • 2021
  • Schisandra nigra Max., a dioecious plant native to Jeju Island in Korea, is cultivated on a small scale for fruit production. As fruit-producing female individuals are generally considered to be more valuable than male, early identification of plant sex at the seedling stage is important. In this study, a sequence-characterized amplified region (SCAR) marker associated with a female-specific region in the genome of S. nigra was investigated. Of 120 randomly amplified polymorphic DNA (RAPD) primers, one primer (OPB-03) consistently amplified a 749 bp band in female plants. The female-specific PCR product was isolated and cloned, and the nucleotide sequences were then determined. Southern hybridization performed using the female-specific fragment as a probe produced positive signals only in genomic DNA from the female plants. This result revealed that the 749 bp segment of DNA was present in the genome of female plants but absent in the genome of male plants. A SCAR primer pair was designed based on the RAPD marker to amplify a 436 bp fragment in the genomic DNA of female plants. This primer pair amplified the expected size of DNA fragment in female plants and four monoecious individuals collected from a natural population. The SCAR marker identified in this study can be used to distinguish female-flowering individuals at the seedling stage.

Development of DNA markers linked to resistant gene to Psmodiophora brassicae Woronin in Chinese cabbage (배추무사마귀병 저항성 유전자와 연관된 DNA 마커개발)

  • 한영한;우종규;박철호
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2002.11b
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    • pp.50-50
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    • 2002
  • 배추무사마귀병 저항성 유전 양식을 증명하기 위해서 CR계 F1에서 유래된 F2 세대를 포장시험과 유묘 검정을 실시하였다. F$_2$ 세대의 7 집단은 단인자우성으로 3:1의 분리비를 보였고, 5 집단은 중복 유전자가 관여하는 9:7의 유전 분리비를 보였다. 배추무사마귀병 저항성 유전자와 연관된 DNA 마커를 개발하기 위하여 CR-Saerona F$_2$ 집단을 배추무사마귀병 발병포장에서 재배하여 저항성 평가를 하였다. 220개의 임의의 프라이머를 이용하여 BSA-RAPD (Bulked segregant analysis-Randomly amplified polymorphic DNA)를 수행하였지만 CR-Saerona F2 집단에서 배추무사마귀병 저항성 유전자와 꼭 들어맞는 DNA 마커는 발견되지 않았다. 300개의 임의의 프라이머를 이용하여 CR-Saerona에서 유래된 F$_2$ 세대를 QTL 분석하였다. 저항성 정도는 발병지수에 따라 조사되었고 QTL 분석을 위해 one-way ANOVA 테스트를 하였다. 통계분석 결과 두 프라이머(K16-1, L2-2)가 저항성과의 상관관계를 보여 주었으나 유의성은 인정되지 않았다.

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