• 제목/요약/키워드: random amplified polymorphic DNA(RAPD)

검색결과 313건 처리시간 0.025초

Purity Test of Radish Hybrid Seeds Using Randomly amplified Polymorphic DNA Marker

  • Oh, Sei-myoung;Soontae Kwon
    • 생명과학회지
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    • 제11권1호
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    • pp.65-67
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    • 2001
  • In order to develop a rapid and simple method for testing the purity of radish hybrid seeds using a procedure based on the PCR(Polymerase chain reaction), eighty random primers were screened with the genomic DNA extracted from five day old seedlings of inbred parent lines and their F1 hybrids. Two primers, HRM-02 (5'-GAGACCAGAC-3') and HRM-19(5'-TGAGGCGTGT-3'), generate reproducible unique PCR patterns which can identify each parent lines as well as their hybrids. In actual test of randomly selected hybrid seeds using the two marker primers, the purity tested by one primer was exactly same as that of other primer. It suggests that one marker primer selected in this experiment is enough for the purity test of radish hybrid seeds. We demonstrates the use of RAPD(randomly amplified polymorphic DNAs) markers to identify each of inbred parent lines and hybrids by rapid and simple method.

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Distinction between Cold-sensitive and -tolerant Jute by DNA Polymorphisms

  • Hossain, Mohammad Belayat;Awal, Aleya;Rahman, Mohammad Aminur;Haque, Samiul;Khan, Haseena
    • BMB Reports
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    • 제36권5호
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    • pp.427-432
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    • 2003
  • Jute is the principal coarse fiber for commercial production and use in Bangladesh. Therefore, the development of a high-yielding and environmental-stress tolerant jute variety would be beneficial for the agro economy of Bangladesh. Two molecular fingerprinting techniques, random-amplified polymorphic DNA (RAPD) and amplified-fragment length polymorphism (AFLP) were applied on six jute samples. Two of them were cold-sensitive varieties and the remaining four were cold-tolerant accessions. RAPD and AFLP fingerprints were employed to generate polymorphism between the cold-sensitive varieties and cold-tolerant accessions because of their simplicity, and also because there is no available sequence information on jute. RAPD data were obtained by using 30 arbitrary oligonucleotide primers. Five primers were found to give polymorphism between the varieties that were tested. AFLP fingerprints were generated using 25 combinations of selective-amplification primers. Eight primer combinations gave the best results with 93 polymorphic fragments, and they were able to discriminate the two cold-sensitive and four cold-tolerant jute populations. A cluster analysis, based on the RAPD and AFLP fingerprint data, showed the population-specific grouping of individuals. This information could be useful later in marker-aided selection between the cold-sensitive varieties and cold-tolerant jute accessions.

DNA 다형성 분석에 의한 느타리버섯 단포자 분리주의 유전적 변이성 (Genetic Variability of Pleurotus ostreatus Monospore Isolates by Random Amplified Polymorphic DNA Analysis)

  • 송영재;정미정;김범기;노영덕;류진창;유영복
    • 한국균학회지
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    • 제24권3호통권78호
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    • pp.186-205
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    • 1996
  • 식품으로서의 가치가 증가하고 있는 느타리버섯(Pleurotus ostreatus) 이핵체와 이것의 자실체로부터 얻은 35 단포자분리균주의 핵산연쇄중합반응에 의한 DNA 다형성을 분석하여 몇가지 유전형질과의 관계를 검토하였다. 느타리의 300개 단포자분리균주중에서 다시 이차로 분리한 35균주의 균총생장 속도를 분석한 결과 4균주는 모균주와 유사한 경향을 나타내었고 느린 것, 아주 느린 것으로 나눌 수 있었다. 26개의 Primer를 사용한 RAPD 분석에서 모균주와 단포자분리균주간에 형성된 DNA 다형성 밴드는 345개로, 밴드의 크기는 $0.1{\sim}4.0Kb$였다. Primer J(OPA-01)와 W(OPB-04)는 균주간에 많은 차이를 보였고, 36-MI 103균주는 거의 모든 primer에서 다른 균주와 차이를 나타냈다. RAPD 분석에 의한 모균주와 단포자분리주의 유전유사도는 36-MI 103을 제외하고 약 73% 정도의 유전유사성을 보였으며, 36-MI 103 균주와 다른 분리균주간의 유사도는 49%로 낮았다. 모균주 및 단포자 분리균주들의 유전유사도는 균사 생장속도와 다소 상관이 있는 4개 군으로 나눌 수 있었는데 I군은 이핵성 모균주, II군은 빠른 생장의 단핵주, III군은 중간 및 느린 생장속도의 단핵주, IV군은 아주 느린 생장속도의 단핵주였다. 그러나 교배형과 유연관계는 상관관계가 없는 것으로 나타났다.

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Genetic Linkage Mapping of RAPD Markers Segregating in Korean Ogol Chicken - White Leghorn Backcross Population

  • Hwang, K.C.;Song, K.D.;Kim, T.H.;Jeong, D.K.;Sohn, S.H.;Lillehoj, H.S.;Han, J.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • 제14권3호
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    • pp.302-306
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    • 2001
  • This study was carried out to construct mapping population and to evaluate the methods involved, including polymorphic DNA marker system and appropriate statistical analysis. As an initial step to establish chicken genome mapping project, White Leghorn (WL) and Korean Ogol chicken (KOC) were used for generating backcross population. From 8 initial parents, total 280 backcross progenies were obtained and 40 were used for genotyping and linkage analysis. For development of novel polymorphic markers for KOC, Random Amplified Polymorphic DNA (RAPD) markers specific for this chicken line were generated. Also included in this study were six microsatellite markers from East Lansing map as reference loci. For segregation analysis, 15 RAPD markers and 6 microsatellites were used to genotype the backcross population. Among the RAPD markers that we developed, 2 pairs of markers were identified to be linked and another 4 RAPD markers showed linkage with microsatellites of known map. In summary, this study showed that our backcross population generated from the mating of KOC to WL serves as a valuable genetic resource for genotyping. Furthermore, RAPD markers are proved to be valuable in linkage mapping analysis.

Genetic Analysis of Haimen Chicken Populations Using Decamer Random Markers

  • Olowofeso, O.;Wang, J.Y.;Zhang, P.;Dai, G.J.;Sheng, H.W.;Wu, R.;Wu, X.
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권11호
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    • pp.1519-1523
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    • 2006
  • Through a screening and selection approach method, decamer random markers were used in a technique called random amplified polymorphic DNA (RAPD) assay with 252 genomic DNAs isolated from four major Haimen chicken populations: Rugao (62), Jiangchun (62), Wan-Nan (63) and Cshiqishi (65). A total of 3-score decamer random primers (S241-S260, S1081-S1100 and S1341-S1360) were employed in the preliminary RAPD-polymerase chain reaction (RAPD-PCR) assay with 50 random template DNA samples from all the populations. Four (6.67%) of the primers that produced obvious polymorphic patterns, interpretable and reproducible bands were selected and used with both the individual DNAs from each population and with pooled DNA samples of the four populations in subsequent analyses. The selected primers produced a total of 131 fragments with molecular size ranging from 835 to 4,972 base pairs (bp) when used with the individual DNAs; 105 (80.15%) of these fragments were polymorphic. With the pooled DNAs, 47 stable and characteristic bands with molecular size ranging from 840 to 4,983 bp, of which 23 (48.94%) polymorphic, were also generated. The band-sharing coefficient (BSC) calculated for the individuals in the population and among populations of bulked samples was between 0.8247 (Rugao) and 0.9500 (Cshiqishi); for pairwise populations, it was between 0.7273 (Rugao vs. Wan-Nan) and 0.9367 (Jiangchun vs. Cshiqishi) chicken populations. Using the BSC for individual and pairwise populations, the Nei's standard genetic distances between the chicken populations were determined and ranged from 0.0043 (Jiangchun vs. Cshiqishi) to 0.1375 (Rugao vs. Cshiqishi). The reconstructed dendrogram linked the Jiangchun and Cshiqishi chickens as closely related populations, followed by Wan-Nan, while the Rugao was the most genetically distant among the populations.

Cytochrome c oxidase subunit 1과 RAPD 분석에 의한 한국 전복속의 계통 연구 (Phylogenetic Study of Genus Haliotis in Korea by Cytochrome c Oxidase Subunit 1 and RAPD Analysis)

  • 서용배;강성철;최성석;이종규;정태혁;임한규;김군도
    • 생명과학회지
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    • 제26권4호
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    • pp.406-413
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    • 2016
  • 전복은 전복속(Haliotis)에 속하며 전 세계적으로 식품산업에서 중요한 복족류 연체동물이다. 우리나라에는 6개종; 북방전복(Haliotis discus hannai), 둥근전복(Haliotis discus discus), 왕전복(Haliotis madaka), 말전복(Haliotis gigantea), 오분자기(Haliotis diversicolor diversicolor), 마대오분자기(Haliotis diversicolor supertexta)가 보고되어 있다. 이 연구에서는 우리나라 해역에 서식하는 중ㆍ대형 전복과 4종인 북방전복, 둥근전복, 왕전복, 말전복의 유전학적 유연관계를 분석하기 위하여 미토콘드리아의 cytochrome c oxidase subunit I (COI) 유전자와 Random Amplified Polymorphic DNA (RAPD) 분석법을 실시 하였다. 본 연구의 결과 COI 유전자 분석과 RAPD 분석을 활용하면 4종의 전복 중 북방전복, 둥근전복, 왕전복을 한 그룹으로 나머지 한 그룹을 말전복으로 구분하는 종 분류는 명확히 구분할 수 있었다. 이러한 결과는 전복 교잡육종을 이용한 수출용 전복 신종자 개발에 있어 주요 대상종인 전복과 4종에 대한 유전적 근연 관계를 규정함으로써 향후 교잡육종 연구의 기초 자료를 제공할 수 있을 것으로 사료된다.

RAPD법을 이용한 고구마 품종간 유연관계 평가 (Evaluation of Genetic Relationship among Sweetpotato Cultivars Using Randomly Amplified Polymorphic DNA (RAPD) Analysis)

  • 이긍표;박권우
    • 원예과학기술지
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    • 제16권1호
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    • pp.18-20
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    • 1998
  • 본 실험은 RAPD (random amplified polymorphic DNA)를 이용하여 국내에서 육성된 13개 품종의 고구마 (Ipomoea batatas)를 대상으로 유연관계분류 및 품종구분 가능성을 탐색하였다. RAPD를 이용하여 고구마 품종을 비가중산술법(UPGMA)으로 3개의 그룹표로 분류할 수 있었는데 그룹 I은 '충승 100호'로, 그룹 II는 '은미', '생미', '수원147호'와 '율미', 그룹 III는 '홍미', '진미', '관동95', '선미', '원미', '신율미', '증미', '풍미'로 나뉘어졌다. RAPD를 이용한 분류 결과는 대체로 육성모부본의 유전자형과 일치함을 나타내고 있고, 상이한 점은 영양계의 변이에 의한 것으로 추측된다, 앞으로 이러한 marker system을 이용하여 육종시 조기에 원하는 형질을 갖는 계통을 선별할 수 있을 것이며 이에 따라 다양한 고구마 품종의 육종프로그램과 품종판별에 유용하게 이용될 수 있을 것으로 사료된다.

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RAPD-PCR(Random Amplified Polymorphic DNA - Polymerase Chain Reaction) 방법을 이용한 Listeria monocytogenes의 검색 (Use of RAPD-PCR(Random Amplified Polymorphic DNA-Polymerase Chain Reaction) Method for a Detection of Pathogenic Listeria monocytogenes)

  • 박범준;신언환
    • 한국식품영양학회지
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    • 제17권3호
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    • pp.254-259
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    • 2004
  • Primer 20가지로 Listeria spp.에 대해 screening을 하여 병원균인 L. monocytogenes를 구별하게 하는 RAPD-PCR의 band pattern을 나타내는 10-mer random primer가 OPG-13이 라는 것을 알았다. OPG-13은 GC%가 70%이므로 계산상으로는 annealing 온도가 34$^{\circ}C$이다. 32~36$^{\circ}C$까지 다섯 가지의 온도로 annealing한 결과 L. monocytogenes만이 갖는 특정한 크기의 band가 역시 34$^{\circ}C$에서 형성됨을 알아냈고, 34$^{\circ}C$를 annealing 온도로 정하였다. Line 1부터 4까지 1. monocytogenes ATCC15313, 19111, 19112, 19113은 2개의 700 bp와 1500 bp band를 형성하였고 그 밖의 Listeria spp.들 Line 6부터 11까지 L. ivanovii ATCC 19119, L. grayi ATCC19120, L. murrayi ATCC25401, L. innocua ATCC33090, L. welshimeri ATCC35897, L. seeligeri ATCC35967은 약 2,000 ~ 2,300 bp크기 한 개의 band를 보여 병원성 균과 비병원성 균이 매우 확실하게 구분되는 band pattern이 나타나는 이러한 결과로 10-mer random primer인 OPG-13을 이용한 RAPD-PCR 방법이 병윈균인 L. monocytogenes를 검색하는데 이용될 수 있음을 확인할 수 있었다.

Aroclor 1254 May Induce Common DNA Effects in Developing Paralichthys olivaceus Embryos and Larvae

  • Min, Eun Young;Kang, Ju Chan
    • Fisheries and Aquatic Sciences
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    • 제17권4호
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    • pp.461-469
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    • 2014
  • Polychlorinated biphenyls (PCBs) are persistent pollutants in aquatic environments, often causing the decline or disappearance of wild populations. In this study, we used a random amplified polymorphic DNA (RAPD) assay to evaluate the effects on the genomic DNA of olive flounder embryo and larval stages of exposure to Aroclor 1254 at concentrations of 1, 5, 10, 20, and $40{\mu}g/L$. We compared RAPD fingerprints of exposed and non-exposed samples. Polymorphisms were revealed as the presence and/or absence of DNA fragments between the two samples. A dose-dependent increase in the number of polymorphic bands was observed with Aroclor 1254 treatment. Also, RAPD profiles of animals exposed to Aroclor 1254 exhibited an increase in the frequency values (FV) compared to the control. A phenogram constructed using neighbor-joining method indicated that genomic template stability in developing embryo and larval stages was significantly affected at ${\geq}5{\mu}g/L$. This study suggested that DNA polymorphisms detected by RAPD analysis could be used as an investigative tool for environmental toxicology and as a useful biomarker in early life stages for the detection of potential genotoxicants.

Construction of Linkage Map Using RAPD and SSR Markers in Soybean (Glycine max)

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    • 한국자원식물학회지
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    • 제10권3호
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    • pp.241-246
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    • 1997
  • Linkage maps based on molecular markers are valuable tools in plant breeding and genetic studies. A population of 76 RI lines from the mating of A3733 and PI437.088 was evaluated with Random Amplified Polymorphic DNA(RAPD) and Simple Sequence Repeats (SSR) markers to create soybean molecular linkage map, 302 RAPD and 21 SSR markers were genetically linked and formed forty linkage groups. These linkage groups spanned a genetic distance of 1,775 cM. The average distance between markers was 5.5 cM.

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