• 제목/요약/키워드: restriction fragment length polymorphism(RFLP)

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황해산 참조기(Pseudosciaena polyactis)의 계군 분석을 위한 분자생물학적 방법 검정 (Preliminary Analysis of Molecular Biological Methods for Stock Identification of Small Yellow Croaker(Pseudosciaena polyactis) in the Yellow Sea)

  • 허회권;황규린;인영철;장정순
    • 한국수산과학회지
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    • 제25권6호
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    • pp.474-484
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    • 1992
  • 황해서식 참조기(Pseudosciaena polyactis)에 대한 계군을 분석하기 위한 연구의 일환으로 우선 황해의 목포 연근해에서 채집된 참조기의 난자와 근육으로부터 mt-DNA와 근육 actin을 추출하였다. 난자의 경우 mt-DNA를 추출한 뒤 제한효소로 처리하여 절편다형현상을 관찰하였으며 근육 actin의 경우 단백질 분해효소(Staphylococcus aureus $V_8$ protease)로 처리하여 N-terminal 절편의 다형현상을 각각 관찰하였다. Mt-DNA의 genome 크기는 약 $\16\pm0.2$ Kb였고 전체 절편수는 37개였으며 사용된 20개의 제한효소 중 8개의 제한효소만이 3개 정도의 절편을 보이므로써 유의성을 관찰할 수 있었다 한편 근육 actin을 Staphylococcus aureus $V_8$단백질 분해효소로 처리하였을 때 N-terminal 부위에서 26 KDa 및 16 KDa의 분자량을 갖는 특이절편을 관찰할 수 있었다.

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한국인 남성 운동선수군에서 Osteocalcin 유전자의 C298T 다형성의 분포와 골밀도와의 관계 (The Distribution of C298T Polymorphism in the Oseteocalcin Gene from Korean Male Athletes and its Association with Bone Mineral Density)

  • 정인근;강병용;김지영;오상덕;하남주
    • 약학회지
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    • 제50권1호
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    • pp.26-32
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    • 2006
  • Osteocalcin is a vitamin K dependent and bone specific protein which plays an important role in the regulation of bone and calcium metabolism. In this study, we evaluated the relationship between the C298T polymorphism in the osteocalcin gene and bone mineral density (BMD) in Korean young men and their interaction with physical activity. BMDs of the femoral neck and lumbar spine were measured using dual energy X-ray absorptiometry, and the C298T polymorphism in the osteocalcin gene determined using polymerase chain reaction (PCR)-HindIII restriction fragment length polymorphism (RFLP) method. We did not observe any significant differences in the femoral neck and lumbar spine BMDs across genotypes of this polymorphism in controls, athletes or combined groups, respectively (P>0.05). Therefore, our data suggest that the C298T polymorphism in the osteocalcin gene is not a suitable genetic marker for the susceptibility to BMD.

정신분열병의 CDH2 유전자 다형성 (The CDH2 Gene Polymorphism in Schizophrenia)

  • 이원석;김미경;정한용;우성일;권영준;김종우;이희제
    • 생물정신의학
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    • 제12권1호
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    • pp.62-67
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    • 2005
  • Objective:There has been increasing evidence that neurodevelopmental dysfunction is involved in the pathophysiology of schizophrenia. Cadherin is known to be one of the important molecules in neurodevelopment. This study was performed to examine the relationship between T816C polymorphism of CDH2 gene and schizophrenia. Methods:Genoytypes of T816C polymorphism of CDH2 gene were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 156 Korea patents with schizophrenia and 170 controls. Results:No difference was found between the patients with schizophrenia and the controls in genotype and allele frequencies of T816C polymorphism of CDH2 gene. Conclusion:The results of this study do not support an association between T816C polymorphism of CDH2 gene and schizophrenia. However, it is necessary to investigate other polymorphic regions of CDH2 in schizophrenia.

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The GSTP1 Ile105Val Polymorphism is not Associated with Susceptibility to Colorectal Cancer

  • Khabaz, Mohamad Nidal
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권6호
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    • pp.2949-2953
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    • 2012
  • The glutathione S transferase (GST) family is a major part of cellular defense mechanisms against endogenous and exogenous substances, many of which have carcinogenic potential. Alteration in the expression level or structure of the glutathione-S-transferase (GST) enzymes may lead to inadequate detoxification of potential carcinogens and consequently contribute to cancer development. A member of the glutathione-S-transferase (GST) family, GSTP1, is an attractive candidate for involvement in susceptibility to carcinogen-associated colorectal cancer. An $Ag{\rightarrow}G$ transition in exon 5 resulting in an Ile105Val amino acid substitution has been identified which alters catalytic efficiency. The present study investigated the possible impact of Ile105Val GSTP1 polymorphism on susceptibility to colorectal cancer. in Jordan We examined 90 tissue samples previously diagnosed with colorectal carcinoma, and 56 non-cancerous colon tissues. DNA was extracted from paraffin embedded tissues and the status of the GSTP1 polymorphism was determined using a polymerase chain reaction restriction fragment length polymorphism (RFLP) method. No statistically significant differences were found between colorectal cancer cases and controls for the GSTP1 Ile/Ile, Ile/Val and Val/Val genotypes. The glutathione S-transferase polymorphism was not associated with risk in colorectal cancer cases in Jordan stratified by age, sex, site, grade or tumor stage. In conclusion, the GSTP1 Ile105Val polymorphism is unlikely to affect the risk of colorectal cancer.

16S rDNA 클론들의 RFLP 비교분석에서 얻어진 Pentachlorophenol의 혐기성 분해에 따른 미생물군집의 변화 (Diversity of Uncultured Microorganisms Associated with the Anaerobic Pentachlorophenol Degradation Estimated by Comparative RELP Analysis of PCR-Amplified 16S rDNA Clones)

  • 성창수;권오섭;박영식
    • 미생물학회지
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    • 제33권2호
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    • pp.149-156
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    • 1997
  • Pentachlorophenol(PCP)가 첨가된 혐기성 무기배지에 혐기성 소화조슬럿지와 쓰레기 매립장의 침출수를 각각 접종한 후, 활성을 보이기 전과 후의 각 시료 내에 존재하는 총유전물질로부터 16S rRNA 유전자를 PCR 증폭하여 이들에 대한 restriction fragment length polymorphism(RFLP) 비교분석과 계통수분석을 실시하였다. 3차에 걸친 RFLP 분석 결과 Ala와 Bld로 명명된 두 가지의 FRLP 유형이 두 가지의 활성시료 모두에서 높은 빈도로 발견되었다. 이들 유형에 속하는 모든 클론들의 5'쪽에 해당되는 180개씩의 염기서열분석을 실시하여 계통수 분석을 실시한 결과, 각각의 유형에 속하는 클론들간에는 높은 상동성을 가지는 것으로 확인되었다. 특히 Bld 유형에서는 78%에 해당되는 클론들이 동일한 염기서열을 가지는 것으로 확인되었다. 이들 Ala와 Bld 유형에 속하는 클론들은 PCP에 대한 분해활성의 결과로서 증식된 유사종의 미생물 군집에서 비롯된 것으로 추정된다. 이 두 가지 유형에 속하는 클론들 중 하나씩의 16S rDNA 전체으 염기서열을 분석한 결과 Ala의 클론은 Clostridium ultunae (Genbank No. Z69293)의 염기서열과 89%의 상동성을 보였으며, Bld의 클론은 Thermobacteroides proteolyticus (Genbank No. X69335)의 것과 97%의 상동성을 가지는 것으로 나타났다.

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Analysis of Phylogenetic Relationships among Medicago Species by Proteins Banding Patterns and RFLP Markers

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    • 한국자원식물학회지
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    • 제10권3호
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    • pp.250-257
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    • 1997
  • The relationship of nine Medicago species belonging to four subgenera were analyzed by using SDS-PAGE and restriction fragment length polymorphism (RELP) methodologies. Sixty-eight bands of alcohol and salt soluble proteins and 85-133 RFLP markers were used to estimate the genetic distance among the species. These species were clustered together at around 0.1 to 0.4 level of distance for both kind of markers, indicating that Medicago species have a large genetic similarity. A combined cluster diagram, at a dissimilarity level of 0.3, differentiated nine species in four groups: group 1, M. littoralis , M. truncatulam, M.scutellata and M. rigidula; group 2, M. sativa ; group 3, M. lupulina ; group 4, M. orbicularis, M. radiata and M. minima. All of them, but except for M. minima. corrensponded to the existing four subgenera of the genus Medicago classified by Lesins and Lesins(1979).The most similar species were M. littoralis and M. trucatula and the most dissimilar one was M. lupulina. In separate cluster diagrams based on RFLP and protein markers, some differences were observed. In the case of RFLP or DNA markers, M. sativa (alfalfa) was distantly clustered with other Medicago species. But in the case of protein markers, M. sativa was closely clustered with M. scutellata, M. littorulis and M. truncatula.

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Cryptosporidium hominis Infection Diagnosed by Real-Time PCR-RFLP

  • Cheun, Hyeng-Il;Kim, Kyungjin;Yoon, Sejoung;Lee, Won-Ja;Park, Woo-Yoon;Sim, Seobo;Yu, Jae-Ran
    • Parasites, Hosts and Diseases
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    • 제51권3호
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    • pp.353-355
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    • 2013
  • There are approximately 20 known species of the genus Cryptosporidium, and among these, 8 infect immunocompetent or immunocompromised humans. C. hominis and C. parvum most commonly infect humans. Differentiating between them is important for evaluating potential sources of infection. We report here the development of a simple and accurate real-time PCR-based restriction fragment length polymorphism (RFLP) method to distinguish between C. parvum and C. hominis. Using the CP2 gene as the target, we found that both Cryptosporidium species yielded 224 bp products. In the subsequent RFLP method using TaqI, 2 bands (99 and 125 bp) specific to C. hominis were detected. Using this method, we detected C. hominis infection in 1 of 21 patients with diarrhea, suggesting that this method could facilitate the detection of C. hominis infections.

가와사키병에서의 TNF-alpha 유전자의 다형성 및 관상동맥 합병증과의 연관성 (Polymorphisms of tumor necrosis factor-alpha promotor gene in Kawasaki disease and relation to the risk of coronary artery lesion)

  • 김세화;윤장원;이영혁;천은정
    • Clinical and Experimental Pediatrics
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    • 제52권4호
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    • pp.476-480
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    • 2009
  • 목 적 : 저자들은 가와사키병 환아에서 TNF-alpha 유전자의 다형성을 조사함으로써 가와사키병과 유전자 다형성의 관련 여부를 알아보고, 또한 관상동맥 병변의 발생과 연관이 있는지를 살펴보려 하였다. 방 법 : 2003년 1월부터 2007년 1월까지 가와사키병 환아 51명과 대조군 48명을 대상으로 TNF-alpha 촉진자의 단일 유전자 다형성을 살펴보았으며, 가와사키병 환아 중 관상동맥 병변이 있는 24명(관상동맥병변군)과 관상동맥 이상이 없었던 27명(정상관상동맥군)에서의 유전자 다형성을 또한 비교하였다. 말초 혈액에서 DNA를 추출하여 TNF-alpha 유전자 -308 부위의 촉진자에 위치한 2개의 단일 염기 서열 G/A에 대한 대립 유전자의 다형성을 Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) 방법으로 분석하였다. 결 과 : 가와사키병 환아군에서의 -308번 TNF-alpha 유전자의 다형성은 51명 중 9명으로 17.6%였고, 정상 대조군에서는 48명 중 3명으로 6.8%로 가와사키병 환아군에서 높았으나 통계학적으로 유의성은 없었다. 가와사키병 환아 중 관상동맥병병군 24명 중 3명인 12.5%에서 유전자 다형성이 있었고 정상관상동맥군은 27명 중 6명인 22.2%로 정상관상동맥군에서 더 높은 빈도 이었으나 통계학적으로 유의하지 않았다. 결 론 : 본 연구에서는 가와사키병 환아의 TNF-alpha의 다형성과 가와사키병의 발병과의 관련성이 통계학적으로 유의한 수준은 아니지만 가와사키병 환아에서 G/A 빈도수가 17.6%으로 대조군에서 6.8% 보다 다소 높게 나타난 결과를 얻었고 앞으로 많은 수의 환아를 대상으로 한다면 유의한 차이가 있을 것으로 생각되므로 향후 대규모의 지속적인 연구가 필요할 것이다.

Molecular Detection and Analysis of Sweet potato feathery motile vims from Root and Leaf Tissues of Cultivated Sweet Potato Plants

  • Ryu, Ki-Hyun;Park, Sun-Hee
    • The Plant Pathology Journal
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    • 제18권1호
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    • pp.12-17
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    • 2002
  • For the molecular detection of Sweet potaio feathery mottle virus (SPFMV) from diseased sweet potato plants, reverse transcription and polymerase chain reaction (RT-PCR) was performed with the use of a set of virus-specific primers to amplify an 816 bp product. The viral coat protein gene was selected for the design of the primers. No PCR product was amplified when Turnip mosaic virus, Potato vims Y or Cucumber mosaic virus were used as template in RT-PCR with the SPFMV-specific primers. The lowest concentration of template viral RNA required for detection was 10 fg. The vim was rapidly detected from total nucleic acids of leaves and roots from the virus-infected sweet potato plants as well as from the purified viral RNA by the RT-PCR. Twenty-four sweet potato samples were selected and analyzed by RT-PCR and restriction fragment length polymorphism (RFLP). RFLP analysis of the PCR products showed three restriction patterns, which resulted in some point mutations suggesting the existence of quasi-species for the vims in the infected sweet potato plants.

Genetic Mapping of Resistant Genes in Brassica pekinensis Against Plasmodiophora brassicae Race 6

  • Lee, Gung-Pyo;Baek, Nam-Kwon;Park, Kuen-Woo
    • The Plant Pathology Journal
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    • 제18권5호
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    • pp.266-270
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    • 2002
  • Inbred lines of Chinese cabbage KU-101 (resistant line against Plasmodiophora brassicae race race 6) and CS-113 (susceptible line) were crossed and their progeny lines F$_1$, BC$_1$F$_1$, F$_2$, and F$_3$ were produced for the construction of the genetic linkage map of R brassicae race 6-resistant Brassica campestris ssp. pekinensis genome. Restriction fragment length polymorphism (RFLP) was applied to compare between parents and their f$_2$ progenies with a total of 192 probes and 5 restriction enzymes. The constructed RFLP map covered 1,104 cM with a mean distance between genetic marker of 8.0 cM, and produced 10 linkage groups having 121 genetic loci. The loci of P. brassicae race 6 (CR6)-resistant Brassica genome were determined by interval mapping of quan-titative trait loci (QTL), which resulted from bioassay using the same race of the fungi in P3 population. Resistant loci were estimated in numbers 1 (Gl) and 3 (G3) linkage groups. In the regression test, Gl had a value of4.8 logarithm of odd (LOD) score, while C3 had values of 4.2-7.2. Given these results, the location of the CR6-resistant loci within the Brassica genome map can now be addressed.