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

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DNA 분석법에 의한 한우고기 판별 (Identification of Hanwoo Meat by DNA Analysis)

  • 오홍록;이창수;상병찬;송광택
    • 농업과학연구
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    • 제33권1호
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    • pp.1-10
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    • 2006
  • 본 연구는 개발된 품종 특이적인 DNA 표지인자를 이용하여 한우와 비한우의 고기를 판별하고자 하였다. 한우와 유럽종 고기소들의 유전적 차이를 Random amplified polymorphic DNA(RAPD) 분석으로 조사한 바, 홀스타인, 헤어포드, 에버딘엥거스, 리모진 및 시멘탈과 같은 한우가 아닌 유럽 종들의 RAPD 패턴들은 동일하였으나, 한우는 이들과 다른 패턴을 보였다. 한우는 유럽종들에서 모두 검출된 특정의 밴드가 발견되지 않았다. 1.4Kbp인 밴드는 한우와 수입소 고기를 판별해 낼 수 있는 유용한 DNA 표지인자로 인정되었다. 실제로, 한우 673 마리, 홀스타인 141 마리의 혈액시료 및 수입육 115의 고기시료를 가지고 시행한 실험에서 그 DNA 표지인자는 비한우에서는 256두 중 245두에서 검출되었으나, 한우에서는 673두 중 644두의 시료에서 검출되지 않아서, 비한우의 검출율은 95%, 한우의 비검출율은 96%를 보였다. 이러한 결과는 그 DNA 표지인자를 이용하여 한우와 비한우 고기를 판별할 수 있음을 시사하였다. 그러나 두 품종 사이의 교잡종은 한우나 비한우의 RAPD 패턴 중에서 한 가지를 무원칙하게 선택하여 나타내고 있으므로, 이러한 DNA 표지인자만으로서는 판별하기가 어려웠다.

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RAPD 분석과 multiplex-PCR을 이용한 석창포 감별용 SCAR 마커 개발 (Development of SCAR Markers for the Authentication of Acori Rhizoma Based on the Analysis of RAPD and Multiplex-PCR)

  • 문병철;지윤의;이영미;천진미;이아영;추병길;김호경
    • 한국약용작물학회지
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    • 제19권3호
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    • pp.162-169
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    • 2011
  • The rhizomes and herbal medicines originating from Acorus gramineus, A. calamus, A. tatarinowii, and A. gramineus var. pusilus, show significant similarity, and the correct identification of species is very difficult. Random Amplified Polymorphic DNA (RAPD) and Sequence Characterized Amplified Region (SCAR) were used to develop a reliable method for identification of these four species. Several distinct SCAR markers were developed from species-specific RAPD amplicons for each species. Furthermore, a useful molecular marker was established for multiplex-PCR, in order to the four species could be distinguished concurrently. These markers allow efficient and rapid identification of closely-related Acorus species and will be useful for standardization of herbal medicines.

팽이버섯의 유전적 변이 (Genetic Variation in Flammulina velutipes)

  • 김종봉;정자인
    • 생명과학회지
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    • 제21권10호
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    • pp.1434-1442
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    • 2011
  • ITS 염기서열과 RAPD를 이용하여 F. velutipes 29개의 팽이버섯 품종 간의 유전적 변이를 분석하였다. ITS 부위에서 720 bp의 염기서열을 확인 하였으나 29개의 팽이품종간에 유의적인 차이가 없었다. RAPD 분석 결과 40개의 random primer 중 다형성을 나타내는 primer는 16개였으며, 그 중 뚜렸한 다형성을 띄는 primer는 OPA-2,4,3,9,10,20 이었다. 이들 29개 품종에서 primer에 의해 증폭된 밴드는 모두 3,030개 였으며, DNA 단편의 크기는 200~2,000 bp 사이에 위치하였다. 또한 3,030개의 scrabble RAPD band들을 marker로 하여 Nei-Li's의 방법을 이용한 비유사도 지수행렬을 조사한 결과 전체 29개 품종의 종내 유전적 변이는 3.3~45%였고, 특히 한국 야생팽이의 종내 유전적 변이도는 17~38.6%로 품종 간 다형성을 확인하였다. RAPD 변이에 기초하여 neighbor-joining tree (NJ) 분석에서는 5개의 cluster로 구분되었으며, 각각의 cluster는 품종, 지역 적 특성을 나타내었다. 본 연구 결과 RAPD와 실험을 통해 확인된 OPA, OPB primer의 경우 미확인 팽이품종들을 검색 하는데 분자 유전적 표지 maker로써 이용 할 수 있는 것으로 생각된다.

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.

Development of a sequence-characterized amplified region (SCAR) marker for female off-season flowering detection in date palm (Phoenix dactylifera L.)

  • Lalita Kethirun;Puangpaka Umpunjun;Ngarmnij Chuenboonngarm;Unchera Viboonjun
    • Journal of Plant Biotechnology
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    • 제50권
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    • pp.190-199
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    • 2023
  • Date palm (Phoenix dactylifera L.: Arecaceae) is a dioecious species where only female trees bear fruits. In their natural state, date palms produce dates once a year. However, in Thailand, some trees were observed to produce dates during the off-season, despite no variations in morphology. The availability of such off-season fruits can significantly increase their market value. Interestingly, most female off-season date palms investigated in this study were obtained through micropropagation. Hence, there is an urgent need for genetic markers to distinguish female offseason flowering plantlets within tissue culture systems. In this study, we aimed to develop random amplification of polymorphic DNA-sequence characterized amplified region (RAPD-SCAR) markers for the identification of female off-season flowering date palms cultivated in Thailand. A total of 160 random decamer primers were employed to screen for specific RAPD markers in off-season flowering male and female populations. Out of these, only one primer, OPN-02, generated distinct genomic DNA patterns in female off-season flowering (FOFdp) individuals compared to female seasonal flowering genotypes. Based on the RAPD-specific sequence, specific SCAR primers denoted as FOFdpF and FOFdpR were developed. These SCAR primers amplified a single 517-bp DNA fragment, predominantly found in off-season flowering populations, with an accuracy rate of 60%. These findings underscore the potential of SCAR marker technology for tracking offseason flowering in date palms. Notably, a BLAST analysis revealed a substantial similarity between the SCAR marker sequence and the transcript variant mRNA from Phoenix dactylifera encoding the SET DOMAIN GROUP 40 protein. In Arabidopsis, this protein is involved in the epigenetic regulation of flowering time. The genetic potential of the off-season flowering traits warrants further elucidation.

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.

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.

Estimation of Genetic Variation of Korean Isolates of Phytophthora capsici by Using Molecular Markers

  • Chee, Hee-Youn;Jee, Hyeong-Jin
    • Mycobiology
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    • 제29권1호
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    • pp.43-47
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    • 2001
  • Genetic diversity of 21 Korean Phytophthora capsici isolates was analyzed by using several molecular markers such as random amplified polymorphic DNA(RAPD), M-13, microsatellite and random amplified microsatellite sequences(RAMS). The overall average similarity coefficient among the isolates was 86% based on the combined data obtained by the molecular markers. No molecular markers were found to be associated with hosts or geographic regions. In addition to RAPD, analysis based on repeated sequences such as $(GTG)_5$, M-13 and RAMS could be used to assess population structure of P. capsici.

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Genetic Relationships of Four Korean Oysters Based on RAPD and Nuclear rDNA ITS Sequence Analyses

  • 김우진;이정호;김경길;김영옥;남보희;공희정;정현택
    • 한국패류학회지
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    • 제25권1호
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    • pp.41-49
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    • 2009
  • Random amplified polymorphic DNA (RAPD) marker and sequence analyses of the internal transcribed spacer (ITS) region of ribosomal DNA were used to assess phylogenetic relationships of four Korean oyster species. The average number of species-specific markers identified from five universal rice primers (URPs) by RAPD-PCR was 1.8 for Crassostrea gigas, 3.2 for C. nippona, 3.6 for C. ariakensis, and 4.6 for Ostrea denselamellosa. The length of the ITS (ITS1-5.8S-ITS2) region ranged from 1,001 to 1,206 bp (ITS1, 426-518 bp; 5.8S, 157 bp; and ITS2, 418-536 bp), while the GC content ranged from 55.5-61.1% (ITS1, 56.8-61.8%; 5.8S, 56-57.3%; and ITS2, 54.1-62.2%). A phylogenetic analysis of the oysters based on our RAPD, ITS1, and ITS2 sequence data revealed a close relationship between C. gigas and C. nippona and a distant relationship between the genera Crassostrea and Ostrea. Our results indicated that RAPD and ITS sequence analysis was a useful tool for the elucidation of phylogenetic relationships and for the selection of species-specific markers in Korean oysters.

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Universal Rice Primer (URP)-RAPD 방법에 의한 어류 종 특이 marker의 동정 (Identification of Potential Species-Specific Marker in Several Fish Species by RAPD Using Universal Rice Primers)

  • 김우진;김경길;이정호;박두원
    • 한국수산과학회지
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    • 제36권3호
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    • pp.317-320
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    • 2003
  • Morphologically similar fish species were subjected to the random amplified polymorphic DNA (RAPD) analysis using universal rice primer (URP). The fish species tested were sea basses (Lateolabrax japonicus and L. maculatus), eels (Anguilla japonica, A. bicolor bicolor, A. rostrata, and A. anguilla), and flounders (Limanda yokohamae and L. herzensteinin). Highly reproducible RAPD patterns were observed with several potential species-specific markers. The results indicate that RAPD technique using URP is useful for distinguishing fish psecies in a rapid manner.