• Title/Summary/Keyword: RAPD PCR

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Intraspecific Relationship Analysis of Safflower (Carthamus tinctorius L.) Lines Collected by RAPD Markers (홍화 수집종의 RAPD에 의한 유연관계 분석)

  • Kim Jae-Chul;Choi Seong-Yong;Shin Dong-Hyun;Kim Se-Jong
    • Korean Journal of Plant Resources
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    • v.19 no.2
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    • pp.336-339
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    • 2006
  • This study was conducted to provide the genetic diversity on Safflower collections and to identify the variations which could be utilized in Safflower breeding. The RAPDs analysis was used to clarify the genetic relationships among 32 Safflower collections. Among 37 primers applied in RAPD analysis, 25 primers that generated appropriate PCR products for identification of the genetic characters in safflower collections were used. Amplified PCR showed the highly reproducible bands at $0.1{\sim}4.0kb$. The number of bands amplified in each primer showed the variations ranging from 1 to 9, with the average of 5.6. A total of 25 bands were identified among twenty-five selected primers and 23 bands (19.2%) showed polymorphism. Based on the similarity value of 0.042 in dendrogram derived from the cluster analysis, the 32 Safflower collections were classified into 6 groups. The two main groups, II and III included 12 collections (38%) and 12 collections (38%), respectively.

Fundamental Study for RAPD-PR Analysis in the Silkworm, Bombyx mori (누에 RAPD-PCR 분석을 위한 기초연구)

  • 황재삼;이진성
    • Journal of Sericultural and Entomological Science
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    • v.38 no.1
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    • pp.7-12
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    • 1996
  • Reproducible the random amplified polymorphic DNAs(RAPDs) patterns were obtained in the two silkworm strains(J111, Galwon) by adjusting concentration optimized of Taq DNA polymerase(one unit), dNTP(200$\mu$M), MgCl2(1.5mM) and template DNA(30ng). In addition, anealing temperature ranging 35$^{\circ}C$ to 42$^{\circ}C$ by the adjusted condition was investigated and fixed at 35$^{\circ}C$ in this study. Variation among individuals and between male and female of Jam 113 strain was not authorized. DNA polymorhpisms among silkworms were authorized by five RAPD markers using OPM04 random primer. Using the primer showing polymorhpims between parents(J111, Galwon) in thirty three individuals, RAPD-PCR for F2 analysis was performed and segregated 3 : 1 in the F2 population. Consequently, RAPDs detected in the parents were obtained as genetic markers, which can be used for construction of genetic map for this industrially particular insect, silkworm Bombyx mori

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Genetic Variation of Coreoleuciscus splendidus Populations from Four Major Rivers in Korea as Assessed by RAPD PCR (RAPD PCR에 의한 4대강 쉬리 Coreoleuciscus splendidus 개체군들의 유전변이 분석)

  • Song, Ha-Yoon;Bang, In-Chul
    • Korean Journal of Ichthyology
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    • v.21 no.2
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    • pp.129-133
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    • 2009
  • Random Amplified Polymorphic DNA (RAPD) analysis was used to investigate the genetic variations of Coreoleuciscus splendidus within and among the West Korea Subdistrict populations (in Han and Geum Rivers) and the South Korea Subdistrict populations (in Seomjin and Nakdong Rivers). Twelve random primers were employed to generate RAPD markers. All primers were produced to identify specific RAPD markers between the West and South Korea Subdistrict populations. Analyses of genetic similarity and distance among the West and South Korea Subdistrict populations of C. splendidus also revealed similar results, with low genetic similarity (0.49~0.53) and high distance value (0.63~0.71). UPGMA dendrogram based on genetic distance was also similar in results. Therefore, the West Korea Subdistrict populations and the South Korea Subdistrict populations vary in genetic structure, and C. splendidus in the South Korea Subdistrict may represent a different species.

Analysis of antibiotic susceptibility of Salmonella Enteritidis isolated from Gyeongnam province and the bacterial genotyping by using RAPD-PCR (경남지역에서 분리한 Salmonella Enteritidis의 항생제 감수성 검사 및 random amplification polymorphic DNA (RAPD)-PCR을 이용한 유전형 분석)

  • Kim, Eun-Gyeong;Kim, Min-Kyung;Kwon, Hyun-Ae;Youn, Do-Kyung;Koo, Jeong-Heon;Park, So-Yeon;Lee, Hui-Geun;Jo, Myeong-Hui;Hah, Do-Yun;Kim, Cheol-Ho;Hwang, Bo-Won;Kim, Sang-Hyun
    • Korean Journal of Veterinary Service
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    • v.41 no.3
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    • pp.149-155
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    • 2018
  • Salmonella Enteritidis (S. Enteritidis) are found in animals, humans, and environment. In addition, S. Enteritidis draws attention to the public health concerns due to carriage of antibiotic resistance traits. For these reasons, the prevalence and antibiotic resistance patterns of S. Enteritidis are significant issues with regard to public health. To address this issues, a total of 24 strains of S. Enteritidis from 164 samples collected from several slaughterhouses in Gyeong-Nam province in order for antibiotic resistance profiles. Subsequently, we characterized the genotyping by random amplification polymorphic DNA (RAPD)-PCR. As a result, very high level of resistance to protein synthesis inhibition antibiotics and most isolates were susceptible to others. Six random primers were used for RAPD-PCR to reveal genotypes of S. Enteritidis isolates. One of the primer, P1245, generated 147 distinct RAPD-PCR fragments ranging from 400~3000 bp. The number of RAPD-PCR products ranged from 4 to 8 for this primer. The RAPD-PCR fragments could be placed these strains into 3 subgroups and 2 classes by UPGMA cluster analysis. Interestingly, several S. Enteritidis that isolated from different slaughterhouses showed same genotype. These results showed only limited genetic variation among the isolates, those were grouped into a few different patterns of antibiotic resistance.

A RAPD-PCR Method for the Rapid Detection of Bacillus cereus

  • Lee, Ji-Yeon;Kwon, Gun-Hee;Park, Jae-Yong;Park, Cheon-Seok;Kwon, Dae-Young;Lim, Jin-Kyu;Kim, Jong-Sang;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.21 no.3
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    • pp.274-276
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    • 2011
  • Distinction of Bacillus cereus from other closely related bacilli is challenging and new efficient methods are continually demanded. From our previous work on RAPD profiles of bacilli, we found a possibility that B. cereus strains could be distinguished from other bacilli. In this work, RAPD-PCR profiles of B. cereus strains were obtained using a 10-mer (S30) as a primer, and a B. cereus specific 0.91-kb band was produced from all tested strains. The RAPD-PCR procedure also successfully detected B. cereus from spiked cheonggukjang when B. cereus cells were present at more than $10^2$/g sample.

Phylogenetic Relationships Using ITS2 Sequence and RAPD-PCR Data from Four Species of Korean Pseudo-nitzschia (Bacillariophyceae) (ITS2 부위의 염기서열 및 RAPC-PCR에 의한 Pseudo-nitzschia 4종의 유연관계)

  • Cho, Eun-Seob;Lee, Young-Sik
    • Journal of Life Science
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    • v.14 no.1
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    • pp.32-37
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    • 2004
  • A portion of ribosomal internal transcribed spacer (ITS) 2 was sequenced from the samples of Pseudo nitzschia (P. deticatissima, P. multiseries, P. pungens and P. subfraudulenta) to investigate the genetic characteristics by measuring tile magnitude of genetic diversity and the degree of similarity coefficient using random amplified polymorphic DNAs (RAPD)-PCR patterns. The phylogenetic trees inferred from the genetic distance analyses showed the placement of P. delicatissima formed a quite long distance from p. P. multiseries, P. pungens, and even P. subfraudulenta. The phylogenetic tree from RAPD patterns showed that P. multiseries and P. pungens had dissimilarity coefficient of 0.31, while P. delicatissima and three species of Pseudo-nitzschia had that of 0.81. It is likely thought that the genetic position of P. delicatissima formed far from P. multiseries, P. punges, and P. subfraudulenta. These results imply that ITS2 region is expected to support a useful molecular characters for recognizing at the species level and for even discriminating P. multiseries from P. pungens. RAPD method also will be used to differentiate the species of Pseudo-nitzschia in a short time.

Characteristics Comparisons of Edwardsiella tarda Isolated from Cultured Olive Flounder and Eel (양식넙치와 뱀장어에서 분리된 Edwardsiella tarda의 특성 비교)

  • Kim, Eunheui
    • Journal of fish pathology
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    • v.34 no.1
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    • pp.31-38
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    • 2021
  • The objective of this study was to determine comparative biochemical characteristics and RAPD (random amplified polymorphic DNA) profiles of 18 strains of Edwardsiella tarda isolated from cultured olive flounder (Paralichthys olivaceus) and eel (Anguilla spp) that showed diseases between 1996 and 2010 in Korea. In terms of biochemical properties, they showed four patterns with differences in citrate degradation and production of H2S and indole. All strains were identified as E. tarda. Characteristics of isolates were not classified according to their host. As a result of PCR with E. tarda-specific primer, EDtT, the same band of about 270 bp was detected in all 18 isolates. However, no specific band was detected in type strains of E. tarda or Edwardsiella ictaluri. As a result of RAPD PCR performed with primer No. 5 and No. 6 of a Ready-To-Go-RAPD kit, the band profile showed clear differences among isolates of olive flounder, isolates of eel, and E. tarda and E. ictaluri type strains. It was possible to organize their characteristics according to the origin of host with possibility to develop tools for pathogen monitoring.

Molecular Discrimination of Dinoflagellates Cochlodinium Polykrikoides Margalef, Gyrodinium Impudicum Fraga et Bravo and Gymnodinium Catenatum Graham using RAPD-PCR Method (RAPD-PCR 방법을 이용한 Cochlodinium polykrikoides Gyrodinium impudicum, Gymnodinium catenatum의 분자생물학적 진단)

  • Cho, Eun-Seob
    • Journal of Life Science
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    • v.13 no.5
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    • pp.651-657
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    • 2003
  • Randomly amplified polymorphic DNA (RAPD) analysis was used to study genetic relationships among C. polykrikoides, G. impudicum and G. catenatum, which possess similar morphological features. Four of 12 primers were selected and 59 amplification products ranged from 0.2 kb to 3.0 kb. The number of polymorphic products in C. polykrikoides, G. impudicum and G. catenatum was 16 (27.1%), 8 (13.5%), and 16 (27.1%), respectively, while 17 were monomorphic. Number of species-specific bounds was 26 (44.0%), 34 (57.6%), 26 (44.0%) in C. polykrikoides, G. impudicum and G. catenatum, respectively. The genetic similarity between C. polykrikoides and G. impudicum/G. catenatum was 0.83, whereas G. impudicum and G. catenatum was 0.78. Our results suggest that C. polykrikoides, G. impudicum and G. catenatum are extremely different on the basis of RAPD analysis, despite similarity based on their morphology. The RAPD technique appears to be efficient in detecting genetic variation in these dinoflagellates.

Genetic Analysis of Somaclones Derived from Different Cytotype Plants of Scilla scilloides Complex using RAPD (PCR-RAPD 기법을 이용한 세포 유형이 다른 무릇 (Scilla scilloides Complex) 체세포클론의 유전적 분석)

  • 오정순;방재욱
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.4
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    • pp.235-240
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    • 1999
  • The usefulness of randomly amplified polymorphic DNA (RAPD) was evaluated to access the genetic variation in somaclones derived from different cytotype plants of Scilla scilloides Complex, AA (2n=16), BB (2n=18) and AABB (2n=34). Three arbitrary decamer primers were successfully used to amplify genomic DNA from the somaclones. DNA polymorphism was observed between cytotypes. The total number of bands in AA, BB and AABB somaclones were 110, 116 and 103, and marker bands examined were 15, 19 and 26, respectively. The diversity of types using PCR in AA, AABB and BB somaclones were 39.2%, 72.3% and 45.7%, respectively. RAPD band patterns suggest that type AA is more stable than type BB and AABB. The frequencies of specific band in AA, BB or AABB somaclones were 0.9%, 4.3% and 4.9%, respectively. The applicability and reliability of RAPD markers for evaluating the somaclones are discussed.

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Genotyping Based on Polymerase Chain Reaction of Enterobacter sakazakii Isolates from Powdered Infant Foods

  • Choi, Suk-Ho;Choi, Jae-Won;Lee, Seung-Bae
    • Food Science and Biotechnology
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    • v.17 no.6
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    • pp.1171-1177
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    • 2008
  • This study was undertaken to classify Enterobacter sakazakii isolates from 13 powdered infant formula products, 25 powdered weaning diet products, and 33 weaning diet ingredients on polymerse chain reaction (PCR) methods. The numbers of the isolates from 1 powdered infant formula product, 7 powdered weaning diet products, and 6 weaning diet ingredients were 1, 14, and 8, respectively. The contaminated ingredients were 1 rice powder, 2 millet powders, 2 vegetable powders, and 1 fruit and vegetable premix. PCR with the primer of repetitive extragenic palindromic element (REP-PCR) and random amplification of polymorphic DNA(RAPD) were effective in discriminating among the isolates, but tRNA-PCR and PCR with the primer of l6S-23S internal transcribed spacer (ITS-PCR) were not. Some of E. sakazakii isolates from vegetable powders, fruit and vegetable premix, and millets powders were classified into the clonal groups based on the DNA patterns in the REP-PCR and RAPD analysis. A close genetic relationship among the isolates from some of the powdered weaning diet products and the rice powder was also detected in the cluster analysis based on the DNA patterns in RAPD.