• 제목/요약/키워드: Amplified Fragment Length Polymorphism

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Diversity of Epiphytic and Acid-tolerant Epiphytic Bacterial Communities on Plant Leaves

  • Joung Pil-Mun;Shin Kwang-Soo;Lim Jong-Soon;Park Seong Joo
    • 한국미생물학회:학술대회논문집
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    • 한국미생물학회 2002년도 추계학술대회
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    • pp.100-105
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    • 2002
  • The diversity of epiphytic bacterial communities on deciduous oak tree (Quercus dentate Thunb.) leaves was examined both in the natural forest area with a clean air and in the industrial estate to assess effects of acidic deposition to the phyllosphere using 16S rDNA sequence data. In addition, acid-tolerant epiphytic bacterial communities were compared. A total of 78 epiphytic and 444 acid-tolerant clones were obtained from clone libraries, resulting in 20 and 17 phylotypes by analysis of restriction fragment length polymorphism (RFLP) for PCR-amplified 16S rDNA products. A low bacterial diversity in both areas was found. As tree leaves grow older, bacterial diversities were slightly increased in the level of subphylum. The community structure of epiphytic bacteria in both areas in April consisted of only two subphyla, $\beta-and\;\gamma-Proteobacteria$. In August two additional subphyla in both areas were found, but the composition was a little different, Acidobacteria and Cytophaga-Flexibacter-Bacteroids (CFB) group in the industrial estate and a -Proteobacteria and CFB group in the natural area, respectively. Acidobacteria could be an indicator of epiphytic bacteria for acidic deposition on plant leaves, whereas a -Proteobacteria be one of epiphytic bacteria that naturally survive on leaves that are not affected by acidic deposition. The acid-tolerant bacterial communities in April were composed of two subphyla, $\gamma-Proteobacteria$ and Low G+C gram-positive bacteria in both areas, and in August a-Proteobacteria was added to the community just in the natural forest area. The direct influence of acidic deposition on the acid-tolerant bacterial phylogenetic composition could not be detected in higher taxonomic levels such as subphylum, but at narrower or finer levels it could be observed by a detection of Xanthomonadales group of $\gamma-Proteobacteria$ just in the industrial estate.

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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.

AFLP analysis to assess genomic stability in Solanum regenerants derived from wild and cultivated species

  • Aversano, Riccardo;Di Dato, Francesco;Di Matteo, Antonio;Frusciante, Luigi;Carputo, Domenico
    • Plant Biotechnology Reports
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    • 제5권3호
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    • pp.265-271
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    • 2011
  • The cultivated potato as well as its tuber-bearing relatives are considered model plants for cell and tissue culture, and therefore for exploiting the genetic variation induced by in vitro culture. The association between molecular stability and tissue culture in different genetic backgrounds and ploidy levels has already been explored. However, it still remains to be ascertained whether somaclonal variation differs between callus-derived chromosome-doubled and undoubled regenerants. Our research aimed at investigating, through amplified fragment length polymorphism (AFLP) markers, the genetic changes in marker-banding patterns of diploid and tetraploid regenerants obtained from one clone each of Solanum bulbocastanum Dunal and S. cardiophyllum Lindl (both 2n = 2x = 24) and tetraploids from cultivated S. tuberosum L. (2n = 4x = 48). Pairwise comparisons between the banding patterns of regenerants and parents allowed detecting considerable changes associated to in vitro culture both at diploid and tetraploid level. The percentages of polymorphic bands between diploid and tetraploid regenerants were, respectively, 57 and 69% in S. bulbocastanum and 58 and 63% in S. cardiophyllum. On average, the frequencies of lost parental fragments in regenerants were significantly higher than novel bands both in S. bulbocastanum (48 vs. 22%) and S. tuberosum (36 vs. 18%) regenerants. By contrast, in S. cardiophyllum, a similar incidence of the two events was detected (32 vs. 29%). Our results revealed that structural changes after tissue culture process strongly affected the genome of the species studied, but diploid and tetraploids regenerated plants responded equally.

서해산 일반 꽃게와 보라색 변이 꽃게의 계측형질 및 AFLP 분석에 의한 집단 구조 (Population Structure of the Blue and Purple Types of the Blue Crab Portunus trituberculatus (Miers) from the West Sea of Korea Based on Morphometric Characteristics and AFLP Analysis)

  • 연인자;송미영;황학진;손명호;김종빈;임양재;김영섭;김근식;방인철
    • 한국수산과학회지
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    • 제41권2호
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    • pp.94-101
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    • 2008
  • In Korean waters, there are two color types (blue and purple) of the blue crab Portunus trituberculatus. The blue type is common, but the ratio of the purple type has increased in landings. To determine whether there were significant morphometric or genetic differences between the blue and purple types, crabs caught from the West Sea of Korea were examined. Based on covariance analysis, there were significant differences in 1 of 10 morphometric characteristics of males between the two types, in none of the ten characteristics for females. Using amplified fragment length polymorphism (AFLP) DNA fingerprinting, no specific AFLP marker was detected for each type. The heterozygosity and genetic diversity were very low. Analyses of pairwise distance, the Fst index, and genetic similarity revealed similar results, with very low genetic differentiation. Therefore, there is no significant difference between blue and purple types of the crab from the West Sea of Korea, and the two types in the West Sea can be managed as one stock.

AFLP 분석에 의한 어름치 Hemibarbus mylodon의 유전 다양성 (Genetic Diversity of Endangered Fish Hemibarbus mylodon (Cyprinidae) Assessed by AFLP)

  • 이윤아;윤영은;남윤권;방인철
    • 한국양식학회지
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    • 제21권3호
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    • pp.196-200
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    • 2008
  • 한국 고유종이며 멸종위기에 처해 있는 어름치 Hemibarbus mylodon 3집단 (북한강, 남한강, 임진강)의 유전적 다양성 및 집단의 유전적 구조를 분석하기 위하여 AFLP분석을 실시하였다. 총 15 primer 쌍을 이용한 3집단의 AFLP 분석에서 795개의 bands가 생성되었으며, 집단 내 다형 band의 출현 빈도는 3개 집단에서 북한강 11.9%, 남한강 11.1%, 임진강 13.4%로 유사하게 나타났고, 평균 유사도는 96.6%로 나타났다. 평균 이형 접합율(0.033-0.040)과 유전적 다양도(0.036-0.043)는 매우 낮은 값을 보였다. 3개의 집단의 Pairwise distance 및 pairwise fixation index 역시 매우 낮은 값을 나타내어 본 연구에서 분석한 어름치 3개 집단은 유전적으로 매우 밀접한 근연관계를 나타내었다. 이러한 결과는 추후 본 종의 복원 및 관리에 필요한 기초자료로 활용될 수 있을 것이다.

Weissella 속 유산균의 빠른 동정을 위한 16S rDNA PCR-RFLP 분석법의 적용 (Application of 16S rDNA PCR-RFLP Analysis for the Rapid Identification of Weissella Species)

  • 이명재;조경희;이종훈
    • 한국미생물·생명공학회지
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    • 제38권4호
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    • pp.455-460
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    • 2010
  • 16S rDNA 특이적 PCR과 증폭산물의 제한효소 처리 후, 나타나는 단편의 크기를 분석하는 PCR-RFLP 분석법을 김치에서 빈번하게 검출되는 Weissella 속 균주 10종의 신속하고 정확한 동정에 적용하였다. Weissella 속 균주 16S rDNA의 특이적 증폭에는 기존에 보고된 PCR primer를 사용하였지만, annealing 온도는 기존의 조건보다 $4^{\circ}C$ 높게 설정한 $65^{\circ}C$에서 PCR을 수행하였다. 증폭산물은 예상크기인 727bp와 일치하였으며, 제한효소 AluI, MseI, BceAI의 처리를 통하여 나타난 단편의 크기는 제한효소 절단위치 분석으로부터 추정한 단편의 크기와 일치하였다. W. kandleri, W. koreensis, W. confusa, W. minor, W. viridescens, W. cibaria, W. soli는 제한효소 AluI과 MseI의 사용으로 구분이 가능하였으며, W. hellenica, W. paramesenteroides의 경우, BceAI을 사용하면 독립적인 구분이 가능하였다. W. thailandensis의 경우, 본 실험에서 사용한 제한효소 AluI, MseI, BceAI에 의해 독립적인 band 양상은 나타나지 않았지만 나머지 9종과의 절단 양상 비교를 통해 구분이 되었으며, 제한효소 MspI을 사용하면 신속하게 동정할 수 있다.

AFLP에 근거한 서남해안 염생식물 4종 개체군의 최소 보존 면적 결정 (Determination on the Minimum Area for Conservation of Four Halophyte Species from the Southwestern Coast of Korea Based on AFLP)

  • 이점숙;임병선;이욱재
    • Journal of Ecology and Environment
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    • 제29권6호
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    • pp.503-509
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    • 2006
  • 서남해안에 자생하고 있는 염생식물 4종, 갈대(Phragmites communis Trin), 칠면초(Suaeda japonica Makino), 갯잔디(Zoysia sinica Hance), 그리고 해홍나물(Suaeda maritima (L.) Dumort)에 대하여 최소 복원 및 보존 면적의 크기를 AFLP (Amplified Fragment Length Polymorphism)기법을 이용한 유전적 다양성 정보에 근거하여 판정하였다. 각 종들의 조사 지역 개체군내에서 유전적 다양성 지수 $\Psi_{ST}$값은 갈대(P. communis)는 0.3856, 칠면초(S. japonica)는 0.1445, 해홍나물(S. maritima)은 0.1669 그리고 갯잔디(Z. sinica) 0.2422을 나타내었다. 칠면초가 0.1445로 가장 낮은 값을 나타내었고 갈대가 0.3856로 가장 높은 값을 나타내었다. 즉, 본 조사 대상 군락 중에서 칠면초 개체군이 가장 높은 유전적 다양성을 나타내었고, 갈대 개체군이 가장 낮은 유전적 다양성을 나타내는 것으로 판단되었다. 한편, 각 개체군별 단위 면적당 유전적 다양성 지수에 근거한 최소 복원 및 보존 면적은 갈대(P. communis)는 $500{\times}500m^2$, 칠면초(S. japonica), 해홍나물(S. maritima), 그리고 갯잔디(Z. sinica)들은 각각 $100\times100m^2$로 판정되었다.

Molecular Identification of Vaginal Lactobacillus spp. Isolated from Korean Women

  • CHANG, CHUNG EUN;SYLVIA I. PAVLOVA;LIN TAO;EUN-KI KIM;SEUNG CHUL KIM;HYUN SHIK YUN;JAE-SEONG SO
    • Journal of Microbiology and Biotechnology
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    • 제12권2호
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    • pp.312-317
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    • 2002
  • Indigenous lactobacilli were isolated from vaginas of Korean women for possible use in ecological treatment of bacterial vaginosis. Vaginal swab samples were obtained from a gynecological clinic and streaked on Rogosa SL agar plates to select the most predominant lactobacilli in each sample. The preliminary identification of the isolates as lactobacilli was based on microscopic observation of Gram-positive rod-shaped cell morphology. The initial characterization was performed on 108 isolates in terms of their cell surface hydrophobicity (CSH), antimicrobial activity, and hydrogen peroxide (H₂O₂) production capability, and 10 isolates were then selected for further molecular identification. For a rapid procedure to identify lactobacilli, polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analyses of the l6S rRNA genes were applied. The 10 selected lactobacilli and 9 different reference strains of Lactobacillus spp. were characterized by PCR-RFLP where the amplified l6S rDNA was digested with 7 different restriction endonucleases prior to analysis. DNA sequencing of the 16S rRNA gene of one particular isolate, KLB 46, that had been identified as L. crispatus by the PCR-RFLP analysis, further confirmed its identity as L. crispatus.

Genetic Diversity of Sweet potato feathery mottle virus from Sweet Potatoes in Korea

  • Kwak, Hae-Ryun;Kim, Mi-Kyeong;Jung, Mi-Nam;Lee, Su-Heon;Park, Jin-Woo;Kim, Kook-Hyung;Ko, Sug-Ju;Choi, Hong-Soo
    • The Plant Pathology Journal
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    • 제23권1호
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    • pp.13-21
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    • 2007
  • Sweet potato feathery mottle virus(SPFMV) is one of the most prevalent viruses infecting sweet potatoes and occurs widely in sweet potato cultivating areas in Korea. To assess their genetic variation, a total of 28 samples infected with SPFMV were subjected to restriction fragment length polymorphism(RFLP) analysis using DNAs amplified by RT-PCR with specific primer sets corresponding to the coat protein(CP) region of the virus. The similarity matrix by UPGMA procedure indicated that 28 samples infected with SPFMV were classified into three groups based on the number and size of DNA fragments by digestion of CP-encoding regions with 7 enzymes including SalI, AluI, EcoRI, HindIII, FokI, Sau3AI, and DraI bands. Four primer combinations out of 5 designed sets were able to differentiate SPFMV and sweet potato virus G infection, suggesting that these specific primers could be used to differentiate inter-groups of SPFMV. Sequence analysis of the CP genes of 17 SPFMV samples were 97-99% and 91-93% identical at the intra-group and inter-groups of SPFMV, respectively. The N-terminal region of the CP is highly variable and examination of the multiple alignments of amino acid sequences revealed two residues(residues 31 and 32) that were consistently different between SPFMV-O and SPFMV-RC.

The variation of insulin like growth factor 2 maker is associated with growth traits in Thai native (Kradon) pigs

  • Kessara Ampaporn;Rattikan Suwannasing;Pitchayanipa Phongphanich;Supanon Tunim;Monchai Duangjinda
    • Animal Bioscience
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    • 제36권9호
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    • pp.1350-1356
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    • 2023
  • Objective: This study was conducted to investigate polymorphisms of the melanocortin-4 receptor (MC4R) and insulin like growth factor 2 (IGF2) genes and to evaluate the growth traits affected by such polymorphisms in Thai native (Kradon) pigs. Methods: Blood samples and productive data from 91 Kradon pigs were collected. DNA was extracted and quantified, the IGF2 and MC4R genes were amplified, and the polymerase chain reaction (PCR) produces were digested using the PCR-restriction fragment length polymorphism (PCR-RFLP) technique. Genotyping was performed, and the association between genotypes and growth traits on the birth and weaning weights were evaluated. Results: The IGF2 intron7 g.162G>C variations in Kradon pigs were found in three genotypes: i) GG, ii) GC, and iii) CC. The GG genotype frequency was the highest followed by the GC and CC genotypes. The frequencies of the G and C alleles were 0.703 and 0.297, respectively. The MC4R genotype was found in only one genotype (GG). The IGF2 gene pattern was not associated with birth weight traits, whereas the IGF2 gene pattern was related to the weaning weight trait in Kradon pigs. Pigs with the CC and GC genotypes had higher weaning weights than ones with the GG genotype (p<0.001). Conclusion: Thai native Kradon pigs with the CC and GC genotypes of the IGF2 gene have higher weaning weights than pigs with the GG genotype.