• 제목/요약/키워드: Random Amplification of Polymorphic DNA

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DNA Profiles Analysed by Polymerase Chain Reaction-Random Amplified Polymorphic DNAs in Shortnecked Clam (Ruditapes philippinarum) Populations

  • Yoon, Jong-Man;Kim, Yong-Ho;Kim, Jong-Yeon
    • 한국양식학회:학술대회논문집
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    • 한국양식학회 2002년도 춘계 한국양식학회 학술대회 발표요지
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    • pp.172-174
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    • 2002
  • Out of 20 primers, 6 generated a total of 1,11 major and minor RAPD bands, producing approximately 4.2 average polymorphic bands pe primer in shortnecked clam (Ruditapes philippinarum) population from Anmyeondo. The bandsharing value altered form 0.15 to 0.74, with the average of 0.5, as calculated by bandsharing analysis. The RAPD profiles obtained with DNAs of two populations from Anmyeondo and Seocheon, respectively, were considerably different (0.20 and 0.51, respectively). The varying degrees of difference among populations may also be of relevance to the retricted hybridization of wild bivalve. Besides gene mapping and breeding applications, PCR-RAPD system could be very useful for the rapid certification and quality control of seed production and for every projects based on PCR amplification of specific bivalve DNA fragments.

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연초에서 발생하는 복숭아혹진딧물(Myzus persicae)형태형 2종의 Random Amplified Polymorphic DNA(RAPD)을 이용한 유전적 유연관계 분석 (Genetic Relationahips of the Two Morphorogical Types of Myzus persicae(Homoptera:Aphididae) Collected from Tobacco Plants Based on Random Amplified Polymorphic DNA(RAPD))

  • 채순용;이기원;김상석;장영덕
    • 한국응용곤충학회지
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    • 제37권1호
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    • pp.31-37
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    • 1998
  • 연초에 채집하여 Blackman(1987)의 기주선호성과 형태적인 특징을 이용하여 분류한 방법에 의해 두가지 타입을 (M. persicae Sulzer와 M. nicotianae Blackman)으로 구분된 무시성충 복숭아 혹진딧물 8 클론의 유전적 특성을 분석을 위하여 RAPD-PCR방법을 이용하였다. 사용된 random primer(10-mer) 100개 중에서 20개의 primer을 선발하였는데 GC content가 70, 80, 90%인 primer에서 각각 26.9%, 50.5% 및 66.^%로 GC content가 높아질수록 PCR결과 band의 양상이 좋게 나타났다. simple matching coefficient를 구하여 matrix를 작성해 본 결과 유사계수(similarity coefficient)의 범위는 0.414~0.808사이이었다. 복숭아혹진딧물 clone 간에 유사계수가 가장 높은 것은 PG2와 PG3 클론으로 0.808로 나타났으며, DBR클론을 기준으로 하여 볼 때 PG2 클론간의 유사계수를 0.414로 유사도가 가장 낮았다. 유사계수를 이용하여 8가지 클론의 진딧물들에 대한 유전적 근연관RP를 살펴보면 M. persicae typer에 속하는 PG1, PG2, PG3 클론들과 M. nicotianae type에 속하는 RED 클론이 유사도 0.643에서 한그룹, M. nicotianae type에 속하는 GR1, GR2, BRN크론들이 유사도 0.636에서 유연관계가 있었으며 그리고 M. persicae type에 the하는 DBR클론 등 세 개의 그룹으로 구분되었다. 따라서 연초에서 발생한 복숭아혹진딧물 형태형 2종 (M. persicae와 M. nicotianae)에 대하여 RAPD 기법을 이용하여 분석해본 결과 뚜렷한 유전적 유연관계는 발견하지 못하였다.

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축산물유래 Listeria monocytogenes의 RAPD typing (Random amplification of polymorphic DNA typing of Listeria monocytogenes isolates from animal products)

  • 이철현;손원근
    • 대한수의학회지
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    • 제49권4호
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    • pp.319-328
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    • 2009
  • This study investigated the epidemiology of Listeria (L.) monocytogenes, a food-borne pathogen. The epidemiology of food-borne pathogens is of great importance for clarifying bacterial origin and preventing bacterial contamination and infection. This work examined 68 L. monocytogenes strains, including 11 reference strains and 57 isolates from imported US beef, domestic meats (beef, pork, chicken meat), raw milk, and milk plants. The random amplified polymorphic DNA (RAPD) techniques were optimized to develop a standard molecular epidemiological analysis of L. monocytogenes. There was great genetic variability among the isolates, which produced 24 and 34 RAPD patterns with primer HLWL85 and HLWL74, respectively. The discriminatory power of the RAPD methods with HLWL85 and HLWL74 primer were very high (DI = 0.957; S ${\geq}$ 80%, S ${\geq}$ 95%). Some RAPD types were specific to origin. A few RAPD types were specific for L. monocytogenes strains belonging to a particular serotype. Using the HLWL85 primer, the strains isolated from milk plants could be distinguished from the other strains. And using the HLWL74 primer, the strains isolated from imported beef (US) could be distinguished completely from the other strains.

해조류로부터 Arbitrary 및 ITS Primer들을 사용한 직접 PCR 유전자 증폭반응의 한계 (Limits of Direct PCR Amplification from Seaweeds Using Arbitrary and ITS Primers)

  • 김용국;진형주;박선미;진덕희;홍용기
    • 생명과학회지
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    • 제9권1호
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    • pp.15-21
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    • 1999
  • PCR 기법을 응용한 RAPD 방법은 대상 생물의 유전정보를 전혀 모르는 상태에도 arbitrary primer를 이용하여 증폭함으로써 생물 종간의 유전적 표식자를 확인할 수 있는 간편하고 유익한 유전분석 방법이다. 이같은 RAPD 방법을 이용하여 해조류 방사무늬 김, 미역, 구멍갈파래에 대하여 DNA를 추출하지 않고 직접 생 엽체 및 생 사상체, 생 배우체를 PCR template로 사용하여 PCR product를 생산하였다. 그러나 specific primer를 이용한 nulear r DNA의 internal trancribed spacer (ITS) 부위는 생성물을 만들지 못하였다. 간편한 LiCl 방법에 의하여 추출된 DNA를 사용하였을 때는 IST 및 RAPD 모두 PCR product를 생산하였다. 방사무늬 김의 엽체 (haploid)와 사상체 (dip-loid)로 부터의 ITS 생성물은 동일하였으나 RAPD 생성물은 사용한 arbitrary primer에 따라 36-50$\%$의 다른 band가 만들어졌다. 또한 직접 생 조직을 사용하였을때와 추출 DNA를 사용하였을 때도 53-57$\%$의 다른 band가 만들어 줬다. 그러므로 RAPD 방법으로 유전분석 실험에는 동일한 ploidy의 조직을 template로서 사용하는 것이 중요하다.

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Rapid molecular authentication of three medicinal plant species, Cynanchum wilfordii, Cynanchum auriculatum, and Polygonum multiflorum (Fallopia multiflorum), by the development of RAPD-derived SCAR markers and multiplex-PCR

  • Moon, Byeong-Cheol;Choo, Byung-Kil;Cheon, Myeong-Sook;Yoon, Tae-Sook;Ji, Yun-Ui;Kim, Bo-Bae;Lee, A-Young;Kim, Ho-Kyoung
    • Plant Biotechnology Reports
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    • 제4권1호
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    • pp.1-7
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    • 2010
  • Definitive identification of original plant species is important for standardizing herbal medicine. The herbal medicines Cynanchi Wilfordii Radix (Baekshuoh in Korean and Beishuwu in Chinese) and Polygoni Multiflori Radix (Hashuoh in Korean and Heshuwu in Chinese) are often misidentified in the Korean herbal market due to morphological similarities and similar names. Therefore, we developed a reliable molecular marker for the identification of Cynanchi Wilfordii Radix and Polygoni Multiflori Radix. We used random amplified polymorphic DNA (RAPD) analysis of three plant species, Polygoni multiflorum, Cynanchum wilfordii, and Cynanchum auriculatum, to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed six sequence characterized amplification region (SCAR) markers for distinguishing Polygoni Multiflori Radix and Cynanchi Wilfordii Radix. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR. These SCAR markers can be used for efficient and rapid authentication of these closely related species, and will be useful for preventing the distribution of adulterants.

Assessment of genetic diversity of Typha angustifolia in the development of cattail stands

  • Min, So-Jung;Kim, Heung-Tae;Kim, Jae-Geun
    • Journal of Ecology and Environment
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    • 제35권1호
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    • pp.27-34
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    • 2012
  • Typha angustifolia has ecological characteristics of clonal growth similar to Phragmites australis. The plant spreads byclonal growth and seed dispersal. In this study, for the three stands which have different settlement age at the Baksilji wetland in Korea, genetic diversity was estimated by random amplification of polymorphic DNA analysis to evaluate the change in genetic diversity of T. angustifolia during stand development in the same population. Stand (ST) 1 was the oldest and ST 4 was the youngest. ST 5 was in a small ditch out of the Baksilji. Although the ST 1, ST 2, and ST 3 did not differ significantly in vegetational or physical environment, the genetic diversity estimated according to Nei's gene diversity (h) and the Shannon index (i) increased in the order of ST 1 < ST 2 < ST 3 contrary to formative age. The genetic diversity of ST 4 was much higher than that of the other three stands. ST 4 has similar abiotic environmental conditions with slight T. angustifolia dominance, and seems to be in the early establishment stage. ST 5 differed from the other stands in vegetational and soil environments, which can result in stressful cattail conditions. Even though the ST 5 stand was not younger than the ST 4 stand, ST 5 showed the highest genetic diversity. Our results indicate that after early settlement of the T. angustifolia population, genetic diversity within the species decreased over time and that the decreasing pattern of genetic diversity within T. angustifolia stands is not likely to occur under stressful conditions.

Construction of a linkage Map in Capsicum annuum L. Using RAPD Markers and Identification of Two QTLs.

  • Yang, Tae-Jin;Kim, Yong-Jae;Park, Hyo-Guen
    • Journal of Plant Biotechnology
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    • 제1권2호
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    • pp.109-115
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    • 1999
  • A linkage map of Capsicum annuum L. was constructed by random amplified polymorphic DNA (RAPD) markers followed in a backcross population of an intraspecific cross between cultivars HDA210 and Yatsufusa. A total of 420 random primers were tested and 311 polymorphic bands were generated by 158 random primers. Among them, 86 Yatsufusa specific bands generated by 52 primers were examined for mapping. Most bands except three segregated in Mendelian fashion fitting the expected 1:1 ratio. The total length of the map was 533 cM distributed in 15 linkage groups. The map distance between adjacent markers ranged 0 to 32.8 cM, with an average distance of 9.1 cM (63 markers). Some markers were clustered and this may be due to the amplification of a repetitive sequence by the RAPDs. Primer pairs for a sequence characterized amplified region (SCAR) were developed and the segregation scores by the SCAR primers were in accordance with the RAPD data. Two QTL markers for number of axillary shoots and for early flowering were developed. One QTL for early flowering located in the linkage group 3 and explained 61 "io of the phenotypic variation. The other QTL for the number of axillary shoots located in the linkage group 4 explained 55 % of the phenotypic variation.tion.

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고온적응성 목이버섯 「현유」 육성 및 특성 (Characteristics and breeding of a thermotolerant ear mushroom, Auricularia auricula-judae 'Hyeonyu')

  • 이기권;유영진
    • 한국버섯학회지
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    • 제15권2호
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    • pp.84-87
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    • 2017
  • 목이버섯의 고온적응성이 우수한 '현유'의 주요특성을 요약하면 다음과 같다. 균사생장적온은 $25^{\circ}C$이고 버섯발생 및 생육온도역시 $26{\sim}36^{\circ}C$로 고온성을 나타내었다. 배양기간은 40~42일, 초발이소요일수는 27일, 생육기간은 15일로 총재배기간은 82~84일이 소요되었다. 갓크기는 56.8 mm이며, 수량은 1 kg배지에서 350 g을 나타내었다. DNA다형성을 분석한 결과 OPA1, OPA7등의 primer에서 개발품종과 중국유래품종 그리고 대조구와 뚜렷한 품종간 밴드차이를 보였다.

누에의 RAPD 분석을 위한 primer의 GC 함량과 사전 제한효소 처리한 주형 DNA의 PCR 증폭효율에 관한 연구 (Studied on Amplificative Efficiency of PCR of Predigested template DNA and GC Contents for RAPD Analysis in the Silkworm, Bombyx mori)

  • 이진성;황재삼;이상몽;황석조;강현아;성승현;서동상
    • 한국응용곤충학회지
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    • 제35권1호
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    • pp.58-65
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    • 1996
  • RAPD-PCR(Random Amplified Polymorphic DNAs-Polymerase Chain Reation) 기법에 누에의 유전적 변이 분석을 위한 첫 단계로 다양한 GC함량을 갖는 random primer에 의해서 증폭되는 DNA 단편의 양상 및 증폭도를 비교하였다. RAPD-PCR을 위한 random primer의 증폭도는 GC함량에 의해서 상당히 영향을 받음이 분석되었다. 특히, 50% GC 함량을 갖는 primer는 그 증폭도에 따라서 4가지의 그룹으로 DNA단편이 증폭되었으며 〔bad amplification (75.5%), poor amplification (11.1%), good and excellent amplification(11.1%)〕, primer의 GC 함량이 증가할수록, 휠씬 더 좋은 증폭도를 보여주었다. 그러나, 40% GC 함량을 갖는 primer에 의해서는 어떤 증폭산물도 관찰되지 않았다. PCR을 수행하기 전에 6가지의 제한효소(BamHI, HindIII, Xbal, HaeIII, MspI, Rsal)를 사용하여 누에 genomic DNA를 처리하여 이를 주형 DNA로 하여 RAPD-PCR을 수행한 결과, 유전적 마커의 생산에 대한 효율이 증가함을 알 수 있었다. 이상의 결과를 종합해 볼 때 60%이상의 GC함량을 갖는 random primer와 전처리한 주형 DNA의 사용은 여러 가지 다른 누에 계통의 동정 및 연관군 지도작성에 따른 경비 및 시간을 줄이는데 효율적이라고 사료된다.

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Monitoring of Gentic Variability in Dicofol-susceptible, Dicofol-resistant, and its Reverse-selected Strains of Tetranychus urticae by RAPD-PCR

  • Song, Cheol;Park, Jin-Hee;Kim, Gil-Hah;Kwon, O-Yu;Cho, Kwang-Yun
    • Journal of Life Science
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    • 제9권1호
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    • pp.14-16
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    • 1999
  • Genetic variability was monitored by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) in dicofol-susceptible (S), dicofol-resistant (R) and its reverse-selected (RS) strains of two-spotted spider mite, of Tetranychus urticae. Before the reverse-selection, RS strain, selected reversely from R strain, was 23-fold resistance ratio at {TEX}$LC_{50}${/TEX} to S strain. The resistance was started to in incline slowly to the resistance level of S strain after one year, and the resistance ratio was 4-fold in the 7 years after then. PCR-amplification of T. urticae DNA showed polymorphism in the amplifications with 12 primers in 100 kinds of arbitrary DNA sequences. RAPD amplification with primer OPR-12 (5`-ACAGGTGCGT-3`) showed amplified bands at 1,000 base pair in the S-and RS-strain, and at 350 base pair in R-strain. The results of polymorphism are genetic variabilities derived from development and selection of resistance in each strain. The peculiarly amplified fragments were guessed to participate in dicofol resistance. From the analysis of genetic similarity, it is inferred the gene composition of S-and RS-strain is much closer than that of R-strain.

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