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

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Molecular characterization of reciprocal crosses of Aerides vandarum and Vanda stangeana (Orchidaceae) at the protocorm stage

  • Kishor, Rajkumar;Devi, H.S.;Jeyaram, K.;Singh, M.R.K.
    • Plant Biotechnology Reports
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    • 제2권2호
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    • pp.145-152
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    • 2008
  • Aerides vandarum and Vanda stangeana are two rare and endangered vandaceous orchids with immense floricultural traits. The intergeneric hybrids were synthesized by performing reciprocal crosses between them. In vitro germination response of the immature hybrid embryos was found to be best on half-strength Murashige and Skoog medium supplemented with 20% (v/v) coconut water/liquid endosperm from tender coconut. Determination of hybridity was made as early as the immature seeds or embryos germinated in vitro, using randomly amplified polymorphic DNA (RAPD) markers. Out of 15 arbitrarily chosen decamer RAPD primers, two were found to be useful in amplification of polymorphic bands specific to the parental species and their presence in the reciprocal crosses. However, a decisive profile that can identify the reciprocal crosses could not be provided by RAPD. Amplification of the trnL-F non-coding regions of chloroplast DNA of the parent species and hybrids aided easy identification of the reciprocal crosses from the fact that maternal inheritance of chloroplast DNA held true for these intergeneric hybrids. Subsequent restriction digestion of the polymerase chain reaction (PCR) amplified trnL-F non-coding regions of chloroplast DNA also consolidated the finding. Such PCR-based molecular markers could be used for early determination of hybridity and easy identification of the reciprocal crosses.

Genetic Polymorphism of Marsh Clam (Corbicula leana) Identified by RAPD- PCR

  • Yoon Jong-Man;Park Kwan-Ha;Choe Sun-Nam
    • Fisheries and Aquatic Sciences
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    • 제6권1호
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    • pp.13-19
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    • 2003
  • Genomic DNA from the muscle of marsh clam (Corbicula leana) from Gochang, Muan and a Chinese site was extracted to identify genetic differences and similarity by randomly amplified polymorphic DNAs-polymerase chain reaction (RAPD- PCR). Out of 20 primers, seven primers produced amplified fragments which were consistently polymorphic. A total of 1,246 amplified products were produced of which 530 were polymorphic $(42.5\%)$. The number of polymorphic bands produced per primer varied from 40 to 122 with an average of 75.7 in marsh clam from Gochang. 3.28 of the 23.0 polymorphic bands per lane were found to be polymorphic. Also, about $4.34\%$ of total polymorphic bands were specific to marsh clam from Gochang. The major common bands of 0.28 kb generated by primer OPB-15 (GGAGGGTGTT) were present in every individuals, which were polymorphic. This common bands in every individuals should be diagnostic of specific strains, species and/or their relatedness. Primer OPB-19 (ACCCCCGAAG) produced the highest number of 12 specific bands. The intra-population variation was revealed in the band patterns identified by this primer. The random primer OPB-12 (CCTTGACGCA) yielded the amplified fragments which were consistently polymorphic between the marsh clams from Gochang and from Muan. This primer produced a total of 77 polymorphic bands: 31 bands from Gochang, 14 from Muan and 32 from the Chinese populations. An average of polymorphic bands were 1.8 from Gochang and 2.5 from the Chinese populations. This value obtained from the Chinese population was higher than those from the two domestic populations. Generally, the RAPD polymorphism generated by these primers may be useful as a genetic marker for strain or population identification of marsh clam.

RAPD 표지인자를 이용한 돌콩 DNA 다형현상 분석 (Identification of DNA polymorphisms in the field bean ( Glycine soza S. and Z. ) using RAPD markers)

  • 이성규
    • 한국초지조사료학회지
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    • 제18권2호
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    • pp.143-150
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    • 1998
  • Six field bean (GI-vcine soza S and Z ) plants were examined for their genetic polymorphisms and intraspecific variations using randomly amplified polymorphic DNA(RAPD) markers. In RAPD analysis of 5 random primers (Rp-1, Rp2, Rp-3, Rp-4, Rp-5), 30 of total 155 bands obtained kom 5 primers were polymorphic and sizes of polymirphic band ranged between 0.5 and 3.0 kb. Number of bands amplyfied per primer was varied from 2 to 11 and average number was 6.0. Genetic variation of intraspecies in the samples of six region was ranged behveen 11 to 25 percent, and genetic similarity among intraspecies was ranged from 0.69 to 0.78. In pairwise genetic similarity test of six field bean plants, Mun and Hoj showed highest coefficient of genetic similarity as 0.67, whereas Sin and Hoj was lowest as 0.45. According to the genetic similarity, the level of intraspecific variation is higher than that of regional distance in GI-vcine soza.

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PCR-RAPD 분석에 의한 붕어(Carassius carassius)의 유전적 유사성 (Genetic Similarity in Crucian Carp(Carassius carassius) by PCR-RAPD Analysis)

  • Yoon, Jong-Man;Kim, Jong-Yeon
    • 한국발생생물학회지:발생과생식
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    • 제5권2호
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    • pp.151-158
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    • 2001
  • 군산지역에 있는 호수와 양식장에서 채집된 붕어(Carassius carassius)의 혈액으로부터 추출된 genomic DNA를 무작위 primer를 이용한 PCR-RAPD 방법에 의해서 유전적 차이를 확인하고자 하였다. 12개 primer 중에서 6개를 이용한 결과 호수산 붕어의 경우 primer 당 약 2.1 polymorphic bands가 나타났고, 총 266개의 높은 RAPD marker가 확인되었으며, 0.18에서 0.76의 bandsharing분석 결과가 나타났다. 군산지역에 있는 호수와 양식장에서 채취된 붕어 2집단간의 RAPD 특징을 bandsharing value로 비교 분석해 본 결과 각각 호수산이 0.47, 양식산이 0.70 으로 나타났으며, 이는 양식산 개체들간에 유사성이 높게 나타났다. 이러한 결과는 군산지역에 있는 양식장의 경우 유사한 환경조건내에서 붕어가 사육되었거나 혹은 오랜 기간동안 근친교배의 결과 이러한 유전적 유사성이 높게 나타난 것으로 사료된다. 달리 말하면 비록 다양한 지리적인 분포가 있더라도 군산지역의 다른 지역으로부터 야생산 붕어 집단의 도입으로 인하여 genomic DNA의 높은 수준의 다양성을 가질 수 있다는 것이다. 일반적으로 primer에 의해서 제시된 RAPD 다형성은 양식대상 어종이면서 온수성 어종인 붕어의 계통 혹은 집단을 확인하기 위한 유전적 표지인자로서 사용될 수 있을 것이다. 그러나 앞으로 집단 및 채집장소의 추가적인 확보 그리고 다른 방법을 통한 연구가 미비한 점을 보완할 수 있는 데 필요하다고 사료된다.

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Selection of RAPD Markers for Phytophthora infestans and PCR Detection of Phytophthora infestans from Potatoes

  • Kim, Kyung-Su;Lee, Youn-Su
    • Journal of Microbiology
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    • 제39권2호
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    • pp.126-132
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    • 2001
  • For rapid and secure differentiation of P. infestans from other Phytophthora species, two fragments obtained from randomly amplified polymorphic DNA (RAPD) profiles were selected as markers. Also, primers for in polymerase chain reaction (PCR) to detect P. infestans specifically were developed by analyzing the sequences of ITSII regions in rDNA of Phytophthora species. The primers, PISP-1 and ITS3 amplified a single. Fragment 450 bp of about in P. infestans, but not in other fungal or bacterial isolates. Annealing temperatures and template DNA quantities were varied for the optimization of PCR conditions. From the result of the PCR detection study, species-specific primers were selected under annealing temperatures ranging from 55$^{\circ}C$ to 61$^{\circ}C$, and template DNA levels ranging from 10 pg to 100 ng.

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Identification of Genetic Markers for Korean Native Cattle (Hanwoo) by RAPD Analysis

  • Yeo Jung Sou;Lee Ji Sun;Lee Chang Hee;Jung Young Ja;Nam Doo Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제5권1호
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    • pp.23-26
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    • 2000
  • In order to develop the specific genetic marker for Korean native cattle (Hanwoo), randomly amplified polymorphic DNA (RAPD) analysis of 6 different cattle breeds was attempted by using 38 decamer primers. In comparison of RAPD patterns, two distinctive DNA bands specific for Hanwoo were detected. One was 296 bp of DNA fragment found to be specific only for female Hanwoo when primer GTCCACACGG was employed. In individual analysis of this RAPD marker was observed only in female individuals with the possibility of $85.3\%$. The other was 521 bp of RAPD marker amplified using TCGGCGATAG and AGCCAGCGAA primers, which showed $83.0\%$ of genetic frequency in 85 male and 68 female individuals tested. Nucleotide sequencing of these genetic markers revealed that 296 bp marker has a short micro satellite-like sequence, ACCACCACAC, and a tandem repeat sequence of microsatellite GAAAAATG in the determined sequence. Two distinctive tandem repeats of microsatellite sequences, MC and GAAGA, were also appeared in 521 bp DNA marker. In BLAST search, any gene having high homology with these markers was not found.

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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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Multi Trait Selection with Restriction for Cutup Carcass Value in Broiler Chicken: Genetic Relatedness of Lines Involved Based on Randomly Amplified Polymorphic DNA

  • Khosravinia, Heshmatollah;Murthy, H.N.N.;Ramesha, K.P.;Govindaiah, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • 제18권11호
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    • pp.1535-1541
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    • 2005
  • Five broiler chicken lines, namely HC, BPB2, CPB2, PB2 and UM1, involving in a selection program and differing in selection intensity and genetic background, were screened for randomly amplified polymorphic DNA (RAPD) polymorphism using 10 selected decamer primers. Nine primers amplified the genomic DNA, generating 200 to 2,500 bp and all detected polymorphism between lines. Out of 74 bands scored using these primers, 34 (50.0%) were found to be polymorphic. The number of polymorphic loci ranged from 3 to 6 with an average of 4.33. Lines differed considerably for within-population genetic similarity estimated by band frequency (WS = 93.55 to 99.25). Between-line genetic similarity estimates based on band sharing as well as on band frequency ranged from 71.35 to 86.45 and from 73.38 to 87.68, respectively. Lines HC and PB2 were the most closely related to the other, while BPB2 and CPB2 appeared to be more distant from each other. The between-line genetic distance based on both band sharing and band frequency revealed the similar trends as for Between-line genetic similarity. Based on BS and BF criteria, BPB2 and CPB2 as well as PB2 and UM1 lines can be merged to launch a new genetic group for further progress in biometrical objectives. A phylogenetic tree, derived using Nei's coefficient of similarity revealed the different pattern of genetic distance between lines.

Genetic variation and relationship of Artemisia capillaris Thunb.(Compositae) by RAPD analysis

  • Kim, Jung-Hyun;Kim, Dong-Kap;Kim, Joo-Hwan
    • 한국자원식물학회지
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    • 제22권3호
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    • pp.242-247
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    • 2009
  • Randomly Amplified Polymorphic DNA (RAPD) was performed to define the genetic variation and relationships of Artemisia capillaris. Fifteen populations by the distributions and habitat were collected to conduct RAPD analysis. RAPD markers were observed mainly between 300bp and 1600bp. Total 72 scorable markers from 7 primers were applied to generate the genetic matrix, and 69 bands were polymorphic and only 3 bands were monomorphic. The genetic dissimilarity matrix by Nei's genetic distance (1972) and UPGMA phenogram were produced from the data matrix. Populations of Artemisia capillaris were clustered with high genetic affinities and cluster patterns were correlated with distributional patterns. Two big groups were clustered as southern area group and middle area group. The closest OTUs were GW2 and GG1 in middle area group, and GB1 from southern area group was clustered with OTUs in middle area group. RAPD data was useful to define the genetic variations and relationships of A. capillaris.