• 제목/요약/키워드: DNA fingerprinting

검색결과 149건 처리시간 0.025초

주요 식중독 그람 음성 세균 4속의 REP-PCR genotyping (REP-PCR Genotyping of Four Major Gram-negative Foodborne Bacterial Pathogens)

  • 정혜진;서현아;김영준;조준일;김근성
    • 한국식품과학회지
    • /
    • 제37권4호
    • /
    • pp.611-617
    • /
    • 2005
  • 본 연구에서는 E. coli. Salmonella, Shigella, Vibrio 등 4속의 주요 식중독유발 그람 음성 세균들을 대상으로 반복성 염기서열인 REP DNA sequence를 응용한 REP-PCR을 실시하였다. 이전의 보고에서 이들 4속의 식중독 유발세균 중 각각 혹은 일부를 대상으로 반복성 염기서열을 이용한 PCR을 적용한 사례는 있지만 그때 적용한 primer, PCR 반응조건 및 전기영동조건 등이 다양하였다. 그러므로 본 연구에서는 이와같은 4속의 세균들에 대하여 최적화된 동일한 primer와 PCR 반응조건 및 전기영동조건을 표준조건으로서 적용하였다. 그 결과로서 모든 4속의 식중독 세균 균주마다 REP-PCR 후 생성되는 fingerprinting pattern에서 속마다 1-3개의 공통적이며 독특한 band가 생성되는 것이 확인되어 이러한 pattern을 이용한 속 수준의 분리 동정과 그와 같은 주요 band들 이외의 부수적인 band들을 고려하여 종 수준까지의 분리도 가능함을 확인하였다. 따라서 본 연구를 통하여 반복적 DNA 염기서열을 이용한 REP-PCR이 주요 식중독 세균의 분리 동정 방법으로 사용될 수 있음을 확인하였다. 또한 본 연구를 통하여 얻은 결과는 더 많은 속(genus)의 식중독세균을 대상으로 한 새로운 분리 동정 방법을 확립하기 위하여 사용될 수 있을 것이다.

Conservation of Swertia chirata through direct shoot multiplication from leaf explants

  • Chaudhuri, Rituparna Kundu;Pal, Amita;Jha, Timir Baran
    • Plant Biotechnology Reports
    • /
    • 제2권3호
    • /
    • pp.213-218
    • /
    • 2008
  • Swertia chirata is an endangered gentian species that prefers to grow at higher altitudes. This ethnomedicinal herb is known primarily for its bitter taste caused by the presence of important phytochemicals that are directly associated with human health benefits. Due to a continuous loss of habitat and inherent problems of seed viability and seed germination, alternative strategies for propagation and conservation are urgently required to prevent the possible extinction of this species. We have formulated a reproducible protocol for the rapid propagation and conservation of this plant using leaves taken from in vitro shoot cultures. Direct induction of more than seven shoot buds per explant was achieved for the first time when the explants were placed on MS medium supplemented with $2.22{\mu}M$ N-6-benzyladenine, $11.6{\mu}M$ kinetin, and $0.5{\mu}M$ ${\alpha}-naphthalene$ acetic acid. Direct organogenesis was noted exclusively from the adaxial surface of the basal segments of leaves. Leaves closer to the apical meristem were more responsive than those farther away from the meristem. Plants raised through direct organogenesis were evaluated for their clonal fidelity by chromosomal analysis and DNA fingerprinting. Complete plants were successfully transferred to the field condition and produced viable seeds. Given the enormous potential of this age-old medicinal plant in terms of potential health-benefitting drugs, this protocol can be used for commercial propagation purposes and to initiate future genetic improvement studies.

Genetic Discrimination of Catharanthus roseus Cultivars by Multivariate Analysis of Fourier Transform Infrared Spectroscopy Data

  • Kim, Suk-Weon;Cho, Soo-Hwa;Chung, Hoe-Il;Liu, Jang-R.
    • Journal of Plant Biotechnology
    • /
    • 제34권3호
    • /
    • pp.201-205
    • /
    • 2007
  • To determine whether pattern recognition based on metabolite fingerprinting for whole cell extracts of higher plants is applied to discriminate plants genetically, leaf samples of eight cultivars of Catharanthus roseus were subjected to Fourier transform infrared spectroscopy (FT-IR). FT-IR fingerprint region data were analyzed by principal component analysis (PCA). Major peaks as biomarkers were identified as the most significant contributors to distinguish samples by using genetic programming. A hierarchical dendrogram based on the results from PCA separated the eight cultivars into two major groups in the same manner as the dendrograms based on genetic fingerprinting methods such as RAPD and AFLP. A slight difference between the dendrograms was found only in branching pattern within each subgroup. Therefore, we conclude that the hierarchical dendrogram based on PCA of the FT-IR data represents the most probable chemotaxonomical relationship between cultivars, which is in general agreement with the genetic relationship determined by conventional DNA fingerprinting methods.

Use of Terminal Restriction Length Polymorphism (T-RFLP) Analysis to Evaluate Uncultivable Microbial Community Structure of Soil

  • Chauhan, Puneet Singh;Shagol, Charlotte C.;Yim, Woo-Jong;Tipayno, Sherlyn C.;Kim, Chang-Gi;Sa, Tong-Min
    • 한국토양비료학회지
    • /
    • 제44권1호
    • /
    • pp.127-145
    • /
    • 2011
  • Various environmental ecosystems are valuable sources for microbial ecology studies, and their analyses using recently developed molecular ecological approaches have drawn significant attention within the scientific community. Changes in the microbial community structures due to various anthropogenic activities can be evaluated by various culture-independent methods e.g. ARISA, DGGE, SSCP, T-RFLP, clone library, pyrosequencing, etc. Direct amplification of total community DNA and amplification of most conserved region (16S rRNA) are common initial steps, followed by either fingerprinting or sequencing analysis. Fingerprinting methods are relatively quicker than sequencing analysis in evaluating the changes in the microbial community. Being an efficient, sensitive and time- and cost effective method, T-RFLP is regularly used by many researchers to access the microbial diversity. Among various fingerprinting methods T-RFLP became an important tool in studying the microbial community structure because of its sensitivity and reproducibility. In this present review, we will discuss the important developments in T-RFLP methodology to distinguish the total microbial diversity and community composition in the various ecosystems.

균주간 유전체 지문 비교분석에서 유전형질 일치성의 확률적 한계 분석 (Analysis of Probabilistic Limits of Trait Identity in Inter-Strain Comparison of Genomic Fingerprints of Bacteria)

  • 조영근
    • 미생물학회지
    • /
    • 제47권3호
    • /
    • pp.263-267
    • /
    • 2011
  • 유전체 지문 분석법은 세균 균주간의 친연성을 판정하는데 유용하다. 그러나 친연성이 낮은 두 균주의 지문 사이에서 우연히 발생하는 DNA 단편 크기의 일치성은 유전형질의 일치성의 해석에 오차를 유발한다. 본 연구는 임의의 두 유전체 지문에서 우연히 DNA 단편의 크기가 일치할 확률을 정량하여, 유전체 지문에 근거한 친연성 해석의 유의성을 고찰하였다. 유전형질 일치성 없이 단편 크기가 일치할 확률은 관찰되는 단편의 수, 관찰 가능한 전체 단편의 수와 크기가 일치하는 단편의 수로부터 계산될 수 있는 함수로 분석되었다. 유의성에 가장 큰 영향을 미치는 독립 매개변수는 전체 단편의 수였으며, 우연한 공통 단편의 수를 10개 미만으로 유지하기 위해서는 약 200개 이상의 단편이 지문에서 관찰될 수 있어야 하는 것으로 계산되었다.

자연산 3배체 붕어 (Carassius auratus) 클론 집단에 대한 유전학적 동정 (Genetic Identification on Natural Population of Triploid Crucian Carp, Carassius auraus in Korea)

  • 김응오;이종윤;남윤권;노재구;이상윤;김동수
    • 한국수산과학회지
    • /
    • 제35권6호
    • /
    • pp.589-594
    • /
    • 2002
  • 국내에 서식하는 붕어 (Carassius auratus) 3배체 집단을 동정하고 이들을 대상으로 분자생물학적, 세포유전학적 및 형태학적 특징을 분석하였다. 본 자연산 붕어 3배체 집단은 전형적인 3배체의 세포 유전학적 특성들, 즉 이배체에 비해 1.5배의 세포 및 핵의 크기, 3n=150의 염색체수, 그리고 2배체 (3.6pg/cell)에 비해 1.5배 증가된 DNA 함량 (5.4 pg/cell)을 나타내었다. 또한 본 3배 체붕어 집단은 DNA fingerprinting 분석에 의해 유전적인 구성이 동일한 클론 집단인 것으로 판명되었고 클론 3배체 붕어들은 2배체와 비교시 매우 일양한 형태학적 특성을 나타내었으며 모두 암컷인 전암컷 집단이었다. 본 연구의 분석에 사용된 함안 집단의 3배체 붕어 집단 이외에도 논산, 주문진, 속초 및 삼례 붕어 집단을 대상으로 3배체 출현 빈도를 조사한 결과, 모든 지역에서 $80\%$ 이상의 높은 3배체 빈도를 보였다.

Utilization of DNA Marker-Assisted Selection in Korean Native Animals

  • Yeo, Jong-sou;Kim, Jae-Woo;Chang, Tea-Kyung;Pake, Young-Ae;Nam, Doo-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • 제5권2호
    • /
    • pp.71-78
    • /
    • 2000
  • The recent progress od DNA technologies including DNA fingerprinting (DFP) and random amplified DNA polymorphism (RAPD) analysis make it possible to identify the specific genetic trits of animals and to analyze the genetic diversity and relatedness between or withinspecies or populations. Using those techniquse, some efforts to identify and develop the specific DNA markers based on DNA polymorphism, which are related with economic traits for Korean native animals, Hanwoo(Korean native cattle),Korean native pig and Korean native chicken, have been made in Korea for recent a few years. The developed specific DNA markers successfully characterize the Korean native animals as the unique Korean genetic sources, distinctively from other imported breeds. Some of these DNA markers have been related to some important economic traits for domestic animals, for example, growth rate and marbling for Honwoo, growth rate and back fat thinkness fornative pig, and growth rate, agg weight and agg productivity for native chicken. This means that those markers can be used in important marker-assised selection (MAS) of Korean native domestic animals and further contribute to genetically improve and breed them.

  • PDF

DNA Profiling of Leuconostoc citreum Strains in Fermented Foods by Repetitive Element Polymerase Chain Reaction

  • Kaur, Jasmine;Sharma, Anshul;Lee, Sulhee;Park, Young-Seo
    • Journal of Microbiology and Biotechnology
    • /
    • 제27권10호
    • /
    • pp.1778-1782
    • /
    • 2017
  • To identify and discriminate the bacterial species at the subspecific level, rep-PCR is a reliable genomic fingerprinting tool. Fourteen strains of bacteria were isolated from different food sources, identified as Leuconostoc citreum using 16S rRNA gene sequencing, and amplified using rep-primers (REP, ERIC, and $(GTG)_5$). Fingerprinting patterns generated bands in the range of 300-6,000 bp with REP, 150-6,000 bp with ERIC, and 200-1,700 bp with $(GTG)_5$ primers. In UPGMA dendrogram analysis, 14 strains were clustered into three clades (I, II, and III) with all the primers, thus differentiating them at the molecular level. The present study revealed the differentiation of L. citreum strains using rep-PCR.

Fingerprinting of Rice Genomes Using PCR with Arbitrary Primers

  • Park, Kyong-Hee
    • Preventive Nutrition and Food Science
    • /
    • 제3권2호
    • /
    • pp.198-202
    • /
    • 1998
  • The arbitrary primed polymerase chain reaction (AP-PCR) has been used to detect the genetic alternations in the related species. Simple and reproducible fingerprints of complex genomes can be generated using single arbitrary chosen primers and the PCR. The technique was applied to the Oryza species and characterized the relationship among three cultivars of rice species based on theresult of genomic DNA fingerprints. The results indicated that the polymorphism revealed in rice strains and the differences in the PCR product pattern could be represented for each strainis. There was many variationsin the PCR product pattern between cv. Dongin(japonica type)and cv.Hyangdo (indica type), and our chosen AP-primers can ge as markers for strain identification and verfication.

  • PDF