• Title/Summary/Keyword: polymorphic band

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Identification of Different Species and Dultivars of Brassica by SDS-PAGE, Isozyme and Molecular Marker

  • Mukhlesur Rahman Md.;Hirata Yutaka
    • Journal of Plant Biotechnology
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    • v.7 no.1
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    • pp.27-35
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    • 2005
  • Eighty-five different cultivars of Brassica rapa, B. juncea, B. nap us, B. carinata, B. oleracea and hexaploid Brassica collected from Bangladesh, Japan, China and Denmark were analyzed by SDS-PAGE for seed and leaf protein variations, using esterase, acid phosphatase and peroxidase isozyme analysis. Ten polymorphic bands were identified from seed protein however no identifiable polymorphic band was found in the leaf protein. Polymorphic markers clearly distinguished the different Brassica species as well as yellow sarson (YS) and brown seeded (BS) cultivars of B. rapa. The $F_1$ cross between YS and brown seeded cultivars showed the existance of all poly-morphic bands of the respective parents. The Bangla-deshi and Japanese cultivars of B. rapa differed in the amount of seed protein. In the case of isozyme analysis, esterase showed the highest number of polymorphic bands (13) followed by acid phosphatase (9) and peroxidase (5). These polymorphic markers were very effec-tive for classification of all the species studied in this experiment. In parentage tests using isozymes, the hybridity of intra-and-interspecific crosses of almost all the seedlings could be identified from their respective cross combinations. Esterase polymorphism showed a clear differentiation between YS and BS types of B. rapa. In addition, two esterase polymorphic markers were iden ified to differentiate some cultivars of B. juncea. Segregation patterns in these two esterase bands showed a simple Mendelian monohybrid ratio of 3:1 in $F_2$, 1:1 in test cross and 1:0 in back cross progenies. No polymorphic band was identified to distinguish different cultivars of the same species by acid phosphatase or peroxidase. Polymerase Chain Reaction (PCR) was carried out with seed coat color specific marker of B. juncea. The yellow seeded cultivars produced a strong band at 0.5 kb and weak band 1.2 kb. In the addition of these two specific bands, Japanese yellow-seeded cultivars expressed two more weak bands at 1.0 kb and 1.1 kb. Where the brown seeded cultivars generated a single strong band at 1.1 kb. In segregating population, the yellow seed coat color marker segregated at a ratio 15 (brown) : 1 (yellow), indicating the digenic inheritance pattern of the trait.

Genetic Similarity and Difference of Marsh Clam (Corbicula leana) Obtained by RAPD-PCR

  • Yoon, Jong-Man;Park, Hwan-ha;Choe, Sun-Nam
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2002.05a
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    • pp.279-280
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    • 2002
  • Genomic DNA from the muscle of marsh clam (Corbicula leana) from Gochang was extracted in order to identify genetic differences and similarity by randomly amplified polymorphic DNAs-polymerase chain reaction. 3.28 of the 23.0 polymorphic bands per lane were found to be polymorphic in marsh clam. Also, about 4.34% of total polymorphic bands were either specific to marsh clam. The major common bands of 0.28 kb generated by primer OPB-15 (GGAGGGTGTT) were present in every individuals, respectively, which were polymorphic. This common bands which present in every individuals should be diagnostic of specific strains, species and/or their relatedness. Primer OPB-19 (ACCCCCGAAG) produced the highest number of specific bands, which was 12. The specific minor band of 0.07 kb was present in lane 22, which were polymorphic. Especially, only a specific band (1.35 kb) identifying individuals was observed in lane 22.

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Genetic Similarity and Difference of Marsh Clam (Corbicula leana) Obtained by RAPD-PCR

  • Yoon, Jong-Man;Park, Kwan-Ha;Choe, Sun-Nam
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2002.08a
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    • pp.171-172
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    • 2002
  • Genomic DNA from the muscle of marsh clam (Corbicula leana)from Gochang was extrected in order to identify genetic differences and similarity by randomly amplified polymorphic DNAs-polymerase chain reaction. 3.28 of the 23.0 polymorphic bands per lane were found to be polymorphic in marsh clam. Also, about 4.34% of total polymorphic bands were either specific to marsh clam. The major common bands of 0.28 kb generated by primer OPB-15 (GGAGGGTGTT) were present in every individuals, respectively, which were polymorphic. This common bands which present in every individuals should be diagnostic of specific strains, species and-or their relatedness. Primer OPB-19 (ACCCCCGAAG) produced the highest number of specific bands, which was 12. The specific minor band of 0.07 kb was present in lane 22, which were polymorphic. Especially, only a specific band (1.35kg) identifying individuals was observed in lane 22.

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Primers for typing Listeria spp. using Random Amplified Polymorphic DNA (RAPD) ANalysis (Listeria spp.의 RAPD typing을 위한 Primer의 분리력 비교)

  • 임형근;홍종해;박경진;최원상
    • Journal of Food Hygiene and Safety
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    • v.18 no.2
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    • pp.67-72
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    • 2003
  • Random amplified Polymorphic DNA (RAPD) analysis Is based on the amplification of random DNA segment using a single arbitrary primer. Polymorphic DNA patterns identified by this method can be used for typing Listeria monocytogenes. To select the primers for RAPD typing Listeria spp., the performance of 31 primers were compared by analyzing 13 Listeria spp. reference strains. Reproducible electrophoresis patterns were obtained. Among 31 primers, 6 primers (primer 6, HLWL74, UBC155, UBC127, Lis5, Lis11) showed better differentiation, when discrimination index, band clarity, band number, difficulty of band scoring were considered than the others. These primers will be useful far typing Listeria spp. in the future. Currently, we are under investigation for the RAPD typing of contaminated L. monocytogenes for the risk analysis of pork processing plant using these primers.

Primers for Typing Salmonella spp. using Random Amplified Polymorphic DNA (RAPD) Analysis (Salmonella spp.의 RAPD Typing을 위한 Primer의 분리력 비교)

  • Lim, Hyung-Kum;Lee, Kyung-Hee;Hong, Chong-Hae;Park, Gyung-Jin;Choi, Weon-Sang
    • Journal of Food Hygiene and Safety
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    • v.18 no.4
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    • pp.224-228
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    • 2003
  • Random amplified polymorphic DNA (RAPD) analysis is based on the amplification of random DNA segment using a single arbitratrary primer. For typing Salmonella spp., polymorphic DNA patterns identified by this method can be used. To select the primers for RAPD typing Salmonella spp., the performances of 20 primers were compared by analyzing 16 Salmonella spp. reference strains. Reproducible electrophoresis patterns were obtained. Among the 20 primers tested, 4 primers (A, OPG04, OPG10, OPL03) showed better differentiation than the others. At the time discrimination index, band clarity, band number and difficulty of band scoring were considered. These primers will be useful for typing Salmonella spp. in the future. Curretly, we are under investigation for the RAPD typing of contaminated Slmonella spp. for the risk analysis of pork processing plant using the primers.

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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    • v.18 no.11
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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.

PCR Based Detection of Phellinus linteus using Specific Primers Generated from Universal Rice Primer(URP) Derived PCR Polymorphic Band

  • Kang, Hee-Wan;Park, Dong-Suk;Park, Young-Jin;Lee, Byoung-Moo;Cho, Soo-Muk;Kim, Ki-Tae;Seo, Geon-Sik;Go, Seung-Joo
    • Mycobiology
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    • v.30 no.4
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    • pp.202-207
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    • 2002
  • This study was carried out to develop specific primers for PCR detection of Phellinus linteus. Diverse genomes of 15 Phellinus spp. including five Phellinus linteus isolates were fingerprinted by Primer Universal rice primer(URP)1F. The URP-PCR pattern differentiated P. linteus isolates from other phellinus spp. A polymorphic band(2.8 kb), which is unique for P. linteus isolates, was isolated and sequenced. Twenty four-oligonucleotide primer pairs were designed based on information of DNA sequence. The primer set(PLSPF2/PLSPR1) amplified single band(2.2 kb) of expected size with genomic DNA from seven Phellinus linteus, but not with that of other Phellinus species tested. The primers could be used identically in both DNA samples from mycelium and fruit bodies. This specific primers could offer a useful tool for detecting and identifying P. linteus rapidly.

Analysis Genetic Similarity of Gentiana scabra var. buergeri by Randomly Amplified Polymorphic DNA (RAPD를 이용한 용담의 유전적 유사도 분석)

  • Lee, Hae-Kyung;Lee, Mi-Kyung;Moon, Chang-Sik;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
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    • v.4 no.3
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    • pp.224-230
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    • 1996
  • Randomly amplified polymorphic DNA(RAPD) analysis was applied to detect the molecular polymorphisms in Gentiana scabra var. buergeri. A high level of molecular variability was found among wild plants and cultivars. In genetic analysis, eight of twenty primers were selected and 54 amplification products ranged 2. 2 to 0.2 kb were compared. Twenty - nine amplified products showed polymorphic, while five were monomorphic. Twenty of line specific bands were found. In genetic similarity and cluster analysis using PCR products, three wild plants collected from Naejangsan, Daedunsan and Keojedo and one cultivar Seochunjaerae were grouped into one cluster, while cultivar Jinbujaerae and Japanese line separated into another clusters, respectively. The identification of DNA polymorphisms by the RAPD technique will facilitate the selection of the lines from different origin.

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Survey of Proper Primers and Genetic Analysis of Korean Ginseng (Panax ginseng C.A. Meyer) Variants using the RAPD Technique (Randomly Amplified Polymorphic DNA (RAPD) 기술을 이용한 고려인삼의 유전분석을 위한 Primer 선발 및 변종별 비교)

  • 임용표;신최순;이석종;윤영남;조재성
    • Journal of Ginseng Research
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    • v.17 no.2
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    • pp.153-158
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    • 1993
  • The study was carried out for comparison of variants and development of genetic markers using Randomly Amplified Polymorphic D사A (RAPD) analysis method. The ginseng variants used were as follows: Chungkyung-Chong, Hwangskoog-Chong, KG101 selected by the pureline selection method, and 6 kinds of Jakyung-Chong strains Uinjakyung, Jakyung-Chong 81783, Jakyung-Chong 847913, Jaky tong-Chong 79742, Jinjakyung of USSR, and Mimaki of Japan). Four of 10 RAPD primers showed the distinctive polymorphism among 9 ginseng variants and lines, and were selected for more detailed polymorphic analysis. The sequences of 4 selected primers were TGCCGAGCTG (Primer#2), AATCGGGCTG (#4), GAAACGGGTG (U7), and GTGACGTAGG (#8). All primers produced several common bands among the strains. However, when primer # 2 was applied, the electrophoregram showed the specific band at 1.8 kb region in Chungkyung-Chong, Hwangskoog- chong, and KG101, and 1 kb in the Jakyung-Chong 847913. In primer #4, 1.1 kb band was shown in Chungkyung-Chong, Hwangskoog-Chong, KG101, and Jakyung-Chong 79742. In primer # 7, 700 bp band was appeared in Jakyung-Chong 81783 and Jinjakyung of USSR In primer # 8, 800 bp band was observed only in Mimaki, comparing to another strains. When Similarity Index (SI) was calculated, Chungkyung-Chong and Hwngskoog-Chong, and Jakyung- chong 81783 and Jinjakyung of USSR showed the most close SI, 0.11 and 0.08, respectively. The data of KG101, which showed the SI of 0.13 with the group of Chungkyung-Chong and Hwangskoog-Chong, coincided with the fact that it was released from Hwangskoog-Chong by breeding process. The data of Jakyung strains indicated the significant variation among the strains. From these results, RAPD analysis method could be succesively applied to the classification and genetic analysis for breeding of Korean ginseng.

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Random Amplified Polymorphic DNA (RAPD) Identification of Genetic Variation in Chlorella species

  • CHO Jung Jong;KIM Yong-Tae;HUR Sung Bum;KIM Young Tae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.6
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    • pp.761-769
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    • 1996
  • The random amplified polymorphic DNA (RAPD) technique was used to characterize 18 reference strains of microalgae, mostly Chlorella species, collected from various localities around Korea peninsular. Eighteen strains consist of four genera of the family marine Chlorella from 12 samples, two genera of fresh water Chlorella from three samples, and three genera on Nannochloris. Twenty 10-mer anonymous primers were screened for amplification of genomic DNA extracted from samples using the CTAB extraction method. Nineteen of these oligonucleotide primers were positive or band producing. Three of 20 random primers (OPA 10, OPA 12, and OPA 18) resulted in both clear band and a high degree of reproducibility and showed some potential to be used to discriminate individual samples of both genetically hetero-and homogeneous populations, in determining phylogenetic relationships between species within a genus and developing individual fingerprints for each samples.

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