• Title/Summary/Keyword: PCR polymorphic

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Identification of Beauveria spp. Isolated from Mulberry Longicorn Beetle (Apriona germari Hope) using Polymerase Chain Reaction (뽕나무 하늘소(Apriona germari Hope)로부터 Beauveria속 사상균의 분리 및 PCR에 의한 동정)

  • 서종복;진병래
    • Journal of Sericultural and Entomological Science
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    • v.37 no.2
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    • pp.167-171
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    • 1995
  • To develope a microbial pesticide for the control of mulberry longicorn beetle, Apriona germari, Beauveria spp. were isolated from the infected Apriona germari larvae. The morphology of Beauveria spp. was observed by phase contrast and scanning electron microscope. In addition, the Beauveria spp. isolated from Apriona germari were identified by the random amplification of polymorphic DNA using polymerase chain reaction. The results showed that the Beauveria spp., SFB-1A and SFB-3A, isolated from Apriona germari were identified with B. bassiana and B. brongniartii, respectively, suggesting that the random amplification of polymorphic DNA is effective for the identification of Beauveria spp.

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Enhanced Discrimination of Listeria spp. Using RAPD Fingerprinting Complemented by Ribotyping-PCR (리스테리아균의 특성분석을 위한 Molecular Typing 방법의 상호보완)

  • 임형근;홍종해;박경진;최원상
    • Journal of Life Science
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    • v.13 no.5
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    • pp.699-704
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    • 2003
  • The results typed by random amplification of polymorphic DNA (RAPD) were compared with those obtained by Enterobacterial repititive intergenic consensus (ERIC) fingerprinting and ribotyping-PCR. The discriminatory power of RAPD typing was the best among the methods tested. RAPD typing with two different primers for 13 Listeria spp. reference strains produced 11 patterns each. In contrast, ERIC fingerprinting produced 9 patterns and ribotyping-PCR produced 7 patterns each. Composite of two separate RAPD (Lis 11 and primer 6) results or RAPD (Lis11)/ ribotyping-PCR differentiated all 13 Listeria spp. reference strains. Therefore, composite of 2 separate RAPD (Lis11 and primer 6) or composite of RAPD (Lis11)/ribotyping-PCR is expected the most promising approach for typing field isolated Listeria spp. strains.

Genomic Polymorphisms of Genome DNA by Polymerase Chain Reaction-RAPD Analysis Using Arbitrary Primers in Rainbow Trout (PCR-RAPD 기법에 의한 무지개송어 Genome DNA 의 다형현상)

  • Yoon, J.M.
    • Korean Journal of Animal Reproduction
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    • v.23 no.4
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    • pp.303-311
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    • 1999
  • Nuclear DNA was isolated from the sperm cells representing genetic characteristics and genomic polymorphisms of rainbow trout by polymerase chain reaction(PCR) amplification of DNA using arbitrary primers. Genomic DNA fingerprints were generated from rainbow trout sperm DNA by polymerase chain reaction amplification using 20 arbitrary decamers as primers. Out of these primers, 4 generated 17 highly reproducible RAPD markers, producing almost six polymorphic bands per primers. Four of 6 primers tested generated amplified fragments which were polymorphic between different individuals. Polymorphic DNA fragments were reproducibly amplified from independent DNA preparations made from individuals. Rainbow trout was distinctly observed 3 specific DNA markers (2. 3, 2.0 and 1.3kb) in bandsharing. Individual fragments generated using the same arbitrary primer, demonstrated that a single primer detected at least three independent genomic polymorphisms in rainbow trout sperm DNA. The RAPD polymorphism generated by this primer may be used as a genetic marker for individual identification The RAPD-PCR technique has been shown to reveal informative polymorphism in many species of fish. The present results demonstrate that RAPD markers are abundant, reproducible and provide a basis for future gene mapping and MAS in these important aquaculture species using RAPD polymorphic markers. It is concluded that RAPD polymorphisms are useful as genetic markers for fish breed differentiation.

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Analysis of Genetic Relatedness in Alternaria species Producing Host Specific Toxins by PCR Polymorphism

  • Kang, Hee-Wan;Lee, Byung-Ryun;Yu, Seung-Hun
    • The Plant Pathology Journal
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    • v.19 no.5
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    • pp.221-226
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    • 2003
  • Twenty universal rice primers (URPs) were used to detect PCR polymorphisms in 25 isolates of six different Alternaria species producing host specific toxins (HST). Eight URPs could be used to reveal PCR polymorphisms of Alternaria isolates at the intra- and inter-species levels. Specific URP-PCR polymorphic bands that are different from those of the other Alternaria spp. were observed on A. gaisen and A. longipes isolates. Unweighted pair-group method with arithmetic mean (UPGMA) cluster analysis using 94 URP polymorphic bands revealed three clustered groups (A. gaisen group, A. mati complex group, and A. logipes group).

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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Genetic Diversity and DNA Polymorphism in Platycodon grandiflorum DC. Collected from East-Asian Area

  • Park, Chun-Geun;Yan, Zhi-Yi;Lee, Sang-Chul;Shon, Tae-Kwon;Park, Hee-Woon;Jin, Dong-Chun
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.2
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    • pp.115-120
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    • 2005
  • Broadening the genetic base of Platycodon grandiflorum DC. cultivar to sustain improvement requires assessment of genetic diversity available in P. grandiflorum DC.. The objective of this study was to analyze the genetic variation, genetic relationship among 48 samples collected from East-Asian Area by means of RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction) markers. From the 18 primers tested, produced total 211 bands with an average of 11.7 bands per primer and obtained 103 polymorphic band with an average of 5.7 bands per primer,s revealed relatively high percentage of polymorphic bands (48.8%). The genetic similarities calculated from RAPD data varied from 0.688 to 0.994 and were clustered to six major groups on a criterion of 0.78 similarity coefficient. The present study has revealed the significant genetic similarity among the samples tested. The analysis of genetic relationships in P. grandiflorum using RAPD-PCR banding data can be useful for the breed improvement.

Detection of Fragment Length Polymorphism of the VNTR Loci D1S80 and D2S123 by PCR Amplification, PAGE and Silver Staining

  • Nam, Hyun-Suk;Kim, Eun-Hee;Yoon, Wan-Hee;Lee, Kong-Joo
    • BMB Reports
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    • v.28 no.4
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    • pp.359-362
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    • 1995
  • The highly polymorphic variable number of tandem repeat (VNTR) loci in the human genome are informative markers for the genetic characterization of individuals in the paternity test and forensic science as well as for the study of human disease. In this study, VNTR loci D1S80 and D2S123 have been amplified by PCR and the amplified length polymorphic alleles were detected with a discontinuous vertical PAGE system and silver staining. For explicit DNA typing, PCR optimization, in which amplification efficiencies are similar over a wide range of allele sizes, non-specific amplifications are minimal, and new longer alleles have high amplification efficiency, has been performed by changing the PCR reaction buffer composition and thermal cycling conditions. It turned out that adding an appropriate amount of Tween 20 and NP40 to the PCR reaction buffer and raising the annealing temperature to $68^{\circ}C$ in thermal cycling made it possible for optimal VNTR loci amplification. A modified PAGE system for VNTR separation was established. Under these conditions, new longer alleles in the 01580 locus were discovered and 025123 pattern changes in colorectal tumors were observed. These technical tips are valuable for detecting various amplified fragment length polymorphisms.

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Genetic Diversity of Paecilomyces japonica and Cordypces militaris Strains by URP-PCR Fingerprinting (URP-PCR핵산지문에 의한 눈꽃동충하초 (Paecilomyces japonica.)와 번데기동충하초(Cordypces militaris) 유전적 다양성분석)

  • Kim, Jong-Kun;Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.39 no.3
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    • pp.180-184
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    • 2011
  • This study was carried out to identify the genetic characteristics among isolates of Paecilomyces spp.and Cordyceps spp. by URP-PCR analysis. Twenty URP (universal rice primer) primers of 20 mer which were designed from repetitive sequence of rice, were used for producing PCR DNA fingerprints of the mushrooms. Of them, 5 URP primers, URP2F, URP2R, URP9F, URP4R, and URP17R amplified genomic DNA of the mushrooms with polymorphic PCR patterns. On isolates of Cordyceps militaris, primers URP1F, URP2R, URP6R and URP17R produced PCR polymorphic bands of 4 types. Isolates of Cordypces sp. that are isolated from different area of Korea were identical to isolate of C. militaris, while other species of Cordypces were different to the PCR profiles. However, the URP primers did not identify the polymorphism of PCR profile on isolates of P. japonica.