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

검색결과 285건 처리시간 0.032초

Biological Nitrogen Removal System의 세균 군집 분석 (Structure of Bacterial Communities in Biological Nitrogen Removal System)

  • 김경미;이상일;이동훈
    • 미생물학회지
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    • 제42권1호
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    • pp.26-33
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    • 2006
  • 생물학적 질소 제거(Biological nitrogen removal; BNR) 시스템의 효율적인 처리 공정을 이재하기 위하여 질산화 반응조 내 세균 군집 구조를 16S rRNA 유전자의 PCR 및 terminal restriction fragment length polymorphism (T-RELP)방법을 이용하여 분석하였다. 본 연구에서 사용한 BNR 시스템은 국내에서 비교적 많이 적용되고 있는 부상여재를 이용한 고도처리 시스템, Nutrient Removal Laboratory 시스템, 반추기법을 이용한 영양염류 처리 Sequencing Batch Reactor (SBR)시스템이었고, 실험 결과 모든 시료에서 암모니아 산화 세균과 $\beta-proteobacteria$에 해당되는 말단 단편을 확인할 수 있었다. 암모니아 산화세균 군집에서 유래된 말단 단편의 염기서열을 분석한 결과 SBR공정에서는 Nitrosomonas와 Nitrosolobus에 속하는 군집 이 우점종임을 확인할 수 있었다. 그러나 다른 두 공정들에서는 $\beta-proteobacteria$에 속하는 미배양 균주와 Cardococcus australiensis와 염기서열 유사도가 높은 군집이 우점하였다. 또한, 암모니아산화 세균군집을 분석한 결과, SBR 공정이 암모니아 산화세균의 농화 배양에 가장 효과적인 것으로 나타났다. 이러한 결과는 각 BNR 시스템에 동일한 폐수가 유입되었음에도 불구하고 서로 다른 세균 군집 구조를 형성하고 있음을 의미한다.

Identification of Ectomycorrhizal Fungi from Pinus densiflora Seedlings at an Abandoned Coal Mining Spoils

  • Park, Sang-Hyeon;Jeong, Hyeon-Suk;Lee, Yoo-Mee;Eom, Ahn-Heum;Lee, Chang-Seok
    • Journal of Ecology and Environment
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    • 제29권2호
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    • pp.143-149
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    • 2006
  • This study was conducted to identify native ectomycorrhizal (ECM) fungi colonizing Pinus densiflora for revegetation of abandoned coal mines in Korea. Seedlings of P. densiflora growing on coal mining spoils of a study site in Samcheok were collected. ECM roots were observed under stereomicroscope and their DNA were extracted from each root tip for a seedling for molecular identification. A PCR primer pair specific to fungi, ITS1F and ITS4, was used to amplify fungal DNA. Restriction enzymes, Alul and Hinfl were used for restriction fragment length polymorphism (RFLP). Combined with RFLP profiles and sequence analysis, total twenty one taxa were identified from the ECM root tips. Basidiomycetous fungi including Thelephoraceae, Pezizales, Laccaria, Pisolithus and Ascomycetous fungi including ericoid mycorrhizal fungi were identified from this study. Results showed that the most frequently found in the study sites was a species in Thelephoraceae. A possible use of ECM fungi identified in this study for the revegetation of abandoned coal mines with P. densiflora was discussed.

Analysis of Fungicide Sensitivity and Genetic Diversity among Colletotrichum Species in Sweet Persimmon

  • Gang, Geun-Hye;Cho, Hyun Ji;Kim, Hye Sun;Kwack, Yong-Bum;Kwak, Youn-Sig
    • The Plant Pathology Journal
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    • 제31권2호
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    • pp.115-122
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    • 2015
  • Anthracnose, caused by Colletotrichum gloeosporioides (C. gloeosporioides; Teleomorph: Glomerella cingulata), is the most destructive disease that affects sweet persimmon production worldwide. However, the biology, ecology, and genetic variations of C. gloeosporioides remain largely unknown. Therefore, in this study, the development of fungicide resistance and genetic diversity among an anthracnose pathogen population with different geographical origins and the exposure of this population to different cultivation strategies were investigated. A total of 150 pathogen isolates were tested in fungicide sensitivity assays. Five of the tested fungicides suppressed mycelial pathogen growth effectively. However, there were significant differences in the sensitivities exhibited by the pathogen isolates examined. Interestingly, the isolates obtained from practical management orchards versus organic cultivation orchards showed no differences in sensitivity to the same fungicide. PCR-restriction fragment length polymorphism (RFLP) analyses were performed to detect internal transcribed spacer regions and the ${\beta}$-tubulin and glutamine synthetase genes of the pathogens examined. Both the glutamine synthetase and ${\beta}$-tubulin genes contained a complex set of polymorphisms. Based on these results, the pathogens isolated from organic cultivation orchards were found to have more diversity than the isolates obtained from the practical management orchards.

Molecular Sexing and Species Identification of the Processed Meat and Sausages of Horse, Cattle and Pig

  • Kim, Yoo-Kyung;Kang, Yong-Jun;Kang, Geun-Ho;Seong, Pil-Nam;Kim, Jin-Hyoung;Park, Beom-Young;Cho, Sang-Rae;Jeong, Dong Kee;Oh, Hong-Shik;Cho, In-Cheol;Han, Sang-Hyun
    • 한국수정란이식학회지
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    • 제31권1호
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    • pp.61-64
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    • 2016
  • We developed a polymerase chain reaction (PCR)-based molecular method for sexing and identification using sexual dimorphism between the Zinc Finger-X and -Y (ZFX-ZFY) gene and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for mitochondrial DNA (mtDNA) cytochrome B (CYTB) gene in meat pieces and commercial sausages from animals of different origins. Sexual dimorphism based on the presence or absence of SINE-like sequence between ZFX and ZFY genes showed distinguishable band patterns between male and female DNA samples and were easily detected by PCR analyses. Male DNA had two PCR products appearing as distinct two bands (ZFX and ZFY), and female DNA had a single band (ZFX). Molecular identification was carried out using PCR-RFLP of CYTB gene, and showed clear species classification results. The results yielded identical information on the sexes and the species of the meat samples collected from providers without any records. The analyses for DNA isolated from commercial sausage showed that pig was the major source but several sausages originated from chicken and Atlantic cod. Applying this PCR-based molecular method was useful and yielded clear sex information and identified the species of various tissue samples originating from livestock.

Development of PCR-RFLP Technique for Identify Several Members of Fusarium incarnatum-equiseti Species Complex and Fusarium fujikuroi Species Complex

  • Pramunadipta, Syafiqa;Widiastuti, Ani;Wibowo, Arif;Suga, Haruhisa;Priyatmojo, Achmadi
    • The Plant Pathology Journal
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    • 제38권3호
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    • pp.254-260
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    • 2022
  • Fusarium incarnatum-equiseti species complex (FIESC) contain over 40 members. The primer pair Smibo1FM/Semi1RM on the RPB2 partial gene has been reported to be able to identify Fusarium semitectum. The F. fujikuroi species complex (FFSC) contains more than 50 members. The F. verticillioides as a member of this complex can be identified by using VER1/VER2 primer pair on the CaM partial gene. In this research, the Smibo1FM/Semi1RM can amplify F. sulawesiense, F. hainanense, F. bubalinum, and F. tanahbumbuense, members of FIESC at 424 bp. The VER1/VER2 can amplify F. verticillioides, F. andiyazi, and F. pseudocircinatum, members of FFSC at 578 bp. Polymerase chain reaction-restriction fragment length polymorphism by using the combination of three restriction enzymes EcoRV, MspI, and HpyAV can differentiate each species of FIESC used. The two restriction enzymes HpaII and NspI can distinguish each species of FFSC used. The proper identification process is required for pathogen control in the field in order to reduce crop yield loss.

Genetic Diversity of Schistosoma haematobium Eggs Isolated from Human Urine in Sudan

  • Quan, Juan-Hua;Choi, In-Wook;Ismail, Hassan Ahmed Hassan Ahmed;Mohamed, Abdoelohab Saed;Jeong, Hoo-Gn;Lee, Jin-Su;Hong, Sung-Tae;Yong, Tai-Soon;Cha, Guang-Ho;Lee, Young-Ha
    • Parasites, Hosts and Diseases
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    • 제53권3호
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    • pp.271-277
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    • 2015
  • The genetic diversity of Schistosoma haematobium remains largely unstudied in comparison to that of Schistosoma mansoni. To characterize the extent of genetic diversity in S. haematobium among its definitive host (humans), we collected S. haematobium eggs from the urine of 73 infected schoolchildren at 5 primary schools in White Nile State, Sudan, and then performed a randomly amplified polymorphic DNA marker ITS2 by PCR-RFLP analysis. Among 73 S. haematobium egg-positive cases, 13 were selected based on the presence of the S. haematobium satellite markers A4 and B2 in their genomic DNA, and used for RFLP analysis. The 13 samples were subjected to an RFLP analysis of the S. haematobium ITS2 region; however, there was no variation in size among the fragments. Compared to the ITS2 sequences obtained for S. haematobium from Kenya, the nucleotide sequences of the ITS2 regions of S. haematobium from 4 areas in Sudan were consistent with those from Kenya (> 99%). In this study, we demonstrate for the first time that most of the S. haematobium population in Sudan consists of a pan-African S. haematobium genotype; however, we also report the discovery of Kenyan strain inflow into White Nile, Sudan.

16S rDNA 염기서열에 의한 청정지역 및 공단지역 내 식물잎권의 내산성세균 군집의 다양성 (Diversity of Acid-Tolerant Epiphytic Bacterial Communities on Plant Leaves in the Industrial Area and the Natural Forest Area Based on 16S rDNA)

  • 정필문;신광수;임종순;이인수;박성주
    • 미생물학회지
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    • 제37권4호
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    • pp.265-272
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    • 2001
  • 깨끗한 대기가 유지되는 청정지역 및 산성강하물의 영향을 받는 공단지역에서 자라는 떡갈나무(Quercus dentate Thunb.) 잎표면에서 분리한 내산성세균 배양으로부터 16S rDNA를 추출하여 모두 444개의 clone을 얻었으며, 이를 대상으로 Restriction fragment length polymorphism (RFLP)을 실시한 결과 총 17종류의 계통형 (phylotype)이 나타났다. 두 지역에서 나타난 대표적인 내산성 잎권세균 군집은 매우 단순하여 ${\gamma}$-Proteobacteria와 low-G+C gram-positive bacteria의 2개 group이었다. 식물 잎의 나이가 들수록 내산성 잎권세균 계통형의 다양성은 현저하게 증가하였다. 내산성 잎권세균 군집 구조는 $\gamma$-Proteobacteria에 속하는 Pseudomonas group과 Enterobacteriaceae, 그리고 low-G+C gram-positive bacteria 즉 Bacillus/Clostridium group에 속하는 Streptococcaceae와 Staphylococcus group이 우점하였다. 산성강하물에 따른 내산성 잎권세균 군집 구조의 변화는 상위 계통분류군(subphylum 수준)에서는 뚜렷이 볼 수 없었지만 보다 하위분류군에서 볼 때 $\gamma$-Proteobacteria의 Xanthomonadales group은 공단지역에서만, 그리고 $\alpha$-Proteobacteria의 Acetobacteraceae는 청정지역에서만 각각 검출되었으므로 이들 세균집단을 산성강서만, 그리고 $\alpha$-Proteobacteria의 Acetobacteraceae는 청정지역에서만 각각 검출되었으므로 이들 세균집단을 산성강 하물에 대한 지표세균으로서 사용할 수 있는 가능성을 남겨 놓았다.

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카스텔라니가시아메바 혹은 대식가시아메바로 분류된 분리주간의 ribosomaIDNA conserved region의 PCR-RFLP의 다양성 (PCR and RFLP variation of conserved region of small subunit ribosomal DNA among Acanthamoeba isolates assigned to either A. castellanii or A. polyphaga)

  • 공현희;정동일
    • Parasites, Hosts and Diseases
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    • 제34권2호
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    • pp.127-134
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    • 1996
  • 형태학적으로 카스텔라니가시아메바 혹은 대식가시아메바로 동정된 12 분리주들과 쿨버트손가시 아메바. 힐리가시아메바(Aconthomoeba hedwi) 팔레스타인가시아메바(A. plestinenis) 별가시아메바의 small subunitribosomal RNA유전자(ssu rDNA) 중 conserved region을 PCRR 증폭하여 제한효노절단부위를 비교하떴다. 별가시아메바의 PCR증폭 산물의 크기는 1.170 bp였고 나머지 분기주들의 것은 910-930 bp 사이였다 카스텔라너가시아메바로 분류건 여섯 주간의 추정 염기치찬율의 평근은 9.BPg였고. 대식가시아떼바로 분류된 분리주간의 그 평근은 9 6U였다. 카스텔 라니가시아메바로 분류된 여섯 분리주들 사이의 최대 염기치환율은 Chang주와 Ma주 사이의 (7 3%)였고 대식가시아메바로 분류된 여섯 분리주간의 최대 염기치환율은 1A/S3주와 KA/S7주 사이의 (16.1%)였다. 이들 종내 최대 염기치환율은 Castellani주 혹은 CCAP 1501/12g주와 KA/S3 주 사이에거 나타난 카스텔라니가시아메바와 대식가시아떼바간의 종간 최소 염기치환율(2.6%)보 다 횔씬 컸나. 쿨버트손가시아메바. 힐리가시아메바 팔레스타인가시아메바 및 별가시아메바의 PCR-RFLP 양상은 카스텔라니가시아메바 또는 대식가시아메바로 동정된 분리주들의 것들과 그리 고 강호간에서도 높은 염기치환율(평균 23 6%)을 보였다. 이상의 성적으로 미루어 보아 가시아메바속의 분류는 재평가 해 보아야 할 건으로 생각된다 A. healyi와 A. palestinensis의 우리말 이름을 각각 힐리가시아메바와 팔레스타인가시아메바로 제안한다.

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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.

Association of Paraoxonase-1(Q192R and L55M) Gene Polymorphisms and Activity with Colorectal Cancer and Effect of Surgical Intervention

  • Ahmed, Nagwa S.;Shafik, Noha M.;Elraheem, Omar Abd;Abou-Elnoeman, Saad-Eldin A.
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권2호
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    • pp.803-809
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    • 2015
  • Background: Colorectal cancer (CRC) is a leading cause of cancer-related death. Oxidative DNA damage may contribute to cancer risk and the antioxidant paraoxonase is one endogenous free radical scavenger in the human body which could therefore exert an influeence. Purpose: Aim of this study was to determine the role of serum arylesterase (ARE) and paraoxonase 1(PON1) activities in CRC patients and to find any association between (PON1) Q192R and L55M gene polymorphisms in CRC patients. Also the serum ARE and PON1 activities in CRC patients will be investigated before and after surgery Materials and Methods: This study involved a total of 50 patients with newly diagnosed CRC and 80 healthy controls. PON1 and ARE activities were determined using an enzymatic spectrophotometric method. PON1 Q192R and L55M gene polymorphisms were determined using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) based restriction fragment analysis. The restriction enzyme AlwI was used to examine the Q192R polymorphism and Hsp92II for the L55M polymorphism. Results: Significant differences in the PON1 Q192R polymorphism were found between patients and controls. The Q allele was more frequent in the patient group than in controls, while the R allele was more frequent in the controls. Significant differences were found in the L55M polymorphism. Additionally, there were significant differences in L and M allele frequencies (p=0.001). The serum activities of PON1 and ARE were low in QQ and MM genotype. Conclusions: serum PON1 and ARE activities were significantly lower in CRC patients compared to healthy subjects. The R allele may protect against colorectal cancer.