• Title/Summary/Keyword: restriction fragment length polymorphism(RFLP)

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Correlation between Selected XRCC2, XRCC3 and RAD51 Gene Polymorphisms and Primary Breast Cancer in Women in Pakistan

  • Qureshi, Z.;Mahjabeen, I.;Baig, R.M.;Kayani, M.A.
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10225-10229
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    • 2015
  • Genetic polymorphisms in homologous recombination repair genes cause an abnormal development of cancerous cells. In the present study we evaluated the possibility of breast cancer association with single nucleotide polymorphisms of RAD51, XRCC2 and XRCC3 genes. Polymorphisms selected in this study were RAD51 135G/C, XRCC2 Arg188His; and XRCC3 Thr241Met. Each polymorphism was genotyped using Polymerase chain reaction-restriction fragment length polymorphism in study cohort of 306 females (156 breast cancer patients and 150 controls). We observed that heterozygous variant genotype (GC) of RAD51 135 G/C polymorphism was associated with a significantly (OR=2.70; 95%CI (0.63-1.79); p<0.03) increased risk of breast cancer. In case of the XRCC3 gene we observed that frequency of heterozygous (OR=2.88; 95%CI (1.02-8.14); p<0.02) and homozygous (OR=1.46; 95%CI (0.89-2.40); p<0.04) genotype of Thr241Met polymorphism were significantly higher in breast cancer patients. For the Arg188His polymorphism of XRCC2, ~2fold increase in breast cancer risk (OR=1.6, 95%CI = 0.73-3.50) was associated with GA genotype with a p value for trend of 0.03. Our results suggest that the 135G/C polymorphism of the RAD51, Thr241Met polymorphism of XRCC3 and Arg188His polymorphism of XRCC2 can be independent markers of breast cancer risk in Pakistan.

Evaluation of the Diversity of Cyclodextrin-Producing Paenibacillus graminis Strains Isolated from Roots and Rhizospheres of Different Plants by Molecular Methods

  • Vollu Renata Estebanez;Fogel Rafael;Santos Silvia Cristina Cunha dos;Mota Fabio Faria da;Seldin Lucy
    • Journal of Microbiology
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    • v.44 no.6
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    • pp.591-599
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    • 2006
  • To address the diversity of cyclodextrin-producing P. graminis strains isolated from wheat roots and rhizospheres of maize and sorghum sown in Australia, Brazil, and France, restriction fragment length polymorphism analysis of part of genes encoding RNA polymerase (rpoB-RFLP) and DNA gyrase subunit B (gyrB-RFLP) was used to produce genetic fingerprints. A phylogenetic tree based on rpoB gene sequences was also constructed. The isolates originated from Brazil could be separated from those from Australia and France, when data from the rpoB-based phylogenetic tree or gyrB-RFLP were considered. These analyses also allowed the separation of all P. graminis strains studied here into four clusters; one group formed by the strains GJK201 and $RSA19^T$, second group formed by the strains MC22.02 and MC04.21, third group formed by the strains TOD61, TOD 221, TOD302, and TOD111, and forth group formed by all strains isolated from plants sown in Cerrado soil, Brazil. As this last group was formed by strains isolated from sorghum and maize sown in the same soil (Cerrado) in Brazil, our results suggest that the diversity of these P. graminis strains is more affected by the soil type than the plant from where they have been isolated.

A Genetic Linkage Map of Soybean with RFLP, RAPD, SSR and Morphological Markers

  • Kim, Hong-Sik;Lee, Suk-Ha;Lee, Yeong-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.45 no.2
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    • pp.123-127
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    • 2000
  • The objective of this study was to develop a linkage map of soybean under the genetic background of Korean soybean. A set of 89 F/sub 5/ lines was developed from a cross between 'Pureunkong', which was released for soy-bean sprout, and 'Jinpumkong 2', which had no beany taste in seed due to lack of lipoxygenase 1, 2, and 3. A linkage map was constructed for this population with a set of 113 genetic markers including 7 restriction fragment length polymorphism (RFLP) markers, 79 randomly amplified polymorphic DNA (RAPD) markers, 24 simple sequence repeat(SSR) markers, and 3 morphological markers. The map defined approximately 807.4 cM of the soybean genome comprising 25 linkage groups with 98 polymorphic markers. Fifteen markers remained unlinked. Seventeen linkage groups identified here could be assigned to the respective 13 linkage groups in the USDA soybean genetic map. RFLP and SSR markers segregated at only single genetic loci. Fourteen of the 25 linkage groups contained at least one SSR marker locus. Map positions of most of the SSR loci and their linkages with RFLP markers were consistent with previous reports of the USDA soybean linkage groups. For RAPD, banding patterns of 13 decamer primers showed independent segregations at two or more marker loci for each primer. Only the segregation at op Y07 locus was expressed with codominant manner among all RAPD loci. As the soybean genetic map in our study is more updated, molecular approaches of agronomically important genes would be useful to improve Korean soybean improvement.

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Effects of Polycyclic Aromatic Hydrocarbons Contamination on the Community Structure of Microorganisms in Gwangyang bay Sediments (다환방향족 탄화수소가 광양만 퇴적토의 미생물 군집구조에 미치는 영향)

  • 권개경;정성영;이정현;현정호;김상진
    • Korean Journal of Environmental Biology
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    • v.22
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    • pp.38-46
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    • 2004
  • Effect of polycyclic aromatic hydrocarbons (PAHs) on the community structure of indigenous microorganisms in Gwangyang Bay sediments was investigated in Mar. & Aug.,2000. Microbial community structure was analyzed using 5'-terminal restriction fragment length polymorphism (T-RFLP) method. Microbial community structure based on T-RFLP method revealed that community differentiated by sampling period except station 1 located near the stream discharge site from Yeosu Industrial Complex. Even, microbial diversity was higher at stations showed relatively high concentrations of PAHs. The microbial community structure was severely changed during the enrichment culture with 1,000 ppm of PAHs mixture. It was also different between cultivated at 8$^{\circ}C$ and 30$^{\circ}C$. The results implied that temperature, poyosity, organic content and etc were more responsible than PAHs on the microbial community structure.

Cultural characteristics and phylogenetic relationships based on RFLP analysis in Pleurotus species

  • Choi, Sun-Gyu;Jang, Kab-Yeul;Kim, Gyu-Hyun;Kong, Won-Sik;Jo, Jae-Sun;Kim, Hae-Yeong;Yoo, Young-Bok
    • Journal of Mushroom
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    • v.12 no.3
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    • pp.145-153
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    • 2014
  • Pleurotus has increased rapidly production and consumption because of highly nutritional value, natural healthy food and so on. The basic studies for Pleurotus need for development of mushroom industry. This study was to investigate the cultural characteristics among 15 strains of 6 species and to analyze their phylogenetic relationships. The cultural characteristics were investigated by mycelial growth activity at different media, temperature and pH. The optimum media for mycelial growth were YM and MCM in most species. The optimum temperature for mycelial growth were $25^{\circ}C$ and $30^{\circ}C$. The optimum pH for mycelial growth were widly range from pH 5.1 to 7.4. Through the RFLP (restriction fragment length polymorphism) of IGS (intergenic spacer) I region in ribosomal DNA, it was analyzed phylogeny of interspecies and intraspecies. Each species was discriminated well as isolates within each species formed clade to be distinguished other species. P. florida was highly similar to P. floridanus, and P. flabellatus was P. cornucopiae. P. fuscus var. ferulae was highly similar to P. eryngii but discriminated different species in analysis of RFLP of IGS I region and showed different characteristics in mycelial culture. RFLP of IGS I region was useful of studying phylogenetic relationships of species and population.

Comparison of PFGE, IS6110-RFLP, and 24-Locus MIRU-VNTR for Molecular Epidemiologic Typing of Mycobacterium tuberculosis Isolates with Known Epidemic Connections

  • Jeon, Semi;Lim, Nara;Park, Sanghee;Park, Misun;Kim, Seonghan
    • Journal of Microbiology and Biotechnology
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    • v.28 no.2
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    • pp.338-346
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    • 2018
  • Two molecular epidemiologic methods, IS6110 restriction fragment length polymorphism (IS6110-RFLP) and 24-locus mycobacterial interspersed repetitive unit-variable-number tandem repeat (MIRU-VNTR), are used worldwide in studies of Mycobacterium tuberculosis (MTB). Conversely, because of its poor resolution, pulsed-field gel electrophoresis (PFGE) is not widely used for MTB. In this study, we improved the 24-locus MIRU-VNTR and PFGE protocols and compared the effectiveness of these approaches for the molecular typing of MTB using 75 clinical isolates obtained from a cohort investigation of high-risk populations infected with MTB. The 24-locus MIRU-VNTR method demonstrated superior discriminatory ability, followed by PFGE and IS6110-RFLP. Next, we analyzed six isolates with clear epidemiologic connections; that is, isolates from patients who attended the same school. IS6110-RFLP and PFGE identified these samples as the same type. By contrast, according to MIRU-VNTR, two isolates differed from four other isolates at one locus each; one isolate was identified as Mtub29 and the other as QUB-26. In summary, the 24-locus MIRU-VNTR assay was the most useful molecular typing method among the three methods investigated due to its discriminatory power, short time required, and availability as an epidemiologic investigation tool. PFGE was the second-best method. Compared with the other loci assessed in the 24-locus MIRU-VNTR assay, the Mtub29 and QUB-26 loci appeared to exhibit greater variability during transmission.

Comparison of community structure of sulfate reducing bacteria in rice paddy and dry farming soils (논과 밭 토양의 황산염 환원세균 군집 구조 비교)

  • Lee, Jung Bae;Park, Kyeong Ryang
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.21-30
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    • 2015
  • The goal of this study was to identify relationships between the composition of sulfate reducing bacterial assemblages and terminal restriction fragment length polymorphism (T-RFLP) patterns in rice paddy and dry farming soils. Samples of organic farming soils, conventional farming soils, and dry field farming soils were collected in August and November. Analyses of the soil chemical composition revealed similar total nitrogen, total carbon and total inorganic phosphorus levels; however, the moisture content and total carbon were higher than in the other soils in both August and November, respectively. Sulfate reducing bacteria utilizing lactic acid were more widely distributed than those that used acetic acid, and the number of sulfate reducing bacteria in organic farming soil was most abundant. Phylogenetic analysis based on 181 clones revealed that most showed low similarity with cultured sulfate reducing bacteria, but more than 90% similarity with an uncultured sulfate reducing bacteria isolated from the environment. T-RFLP analysis revealed that fragments of 91, 357, 395, and 474 bp were most common, and the community structure of sulfate reducing bacteria changed seasonally.

Development of Sequence-Based DNA Markers for Evaluation of Phylogenetic Relationships in Korean Watermelon Varieties

  • Lee, Hee-Jeong;Cho, Hwa-Jin;Lee, Kyung-Ah;Lee, Min-Seon;Shin, Yoon-Seob;Harn, Chee-Hark;Yang, Seung-Gyun;Nahm, Seok-Hyeon
    • Journal of Crop Science and Biotechnology
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    • v.10 no.2
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    • pp.98-105
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    • 2007
  • Phylogenetic relationships in Korean watermelons were evaluated by genetic similarity coefficients using 15 SSR(simple sequence repeat), 14 SCAR(sequence characterized amplified region) and 14 CAPS(sequence characterized amplified region) markers. The SSR markers were selected from previously reported melon and watermelon SSRs through testing polymorphisms within a set of commercial $F_1$ varieties. The SCAR and CAPS markers were developed from polymorphic AFLP(amplified fragment length polymorphism) markers between inbred lines 'BN4001' and 'BN4002'. From the AFLP analysis, 105 polymorphic fragments were identified between the inbred lines using 1,440 primer combinations of EcoRI+CNNN and XbaI+ANNN. Based on the sequencing data of these polymorphic fragments, we synthesized sequence specific primer pairs and detected clear and reliable polymorphisms in 27 primer pairs by indels(insertion/deletion) or RFLP(restriction fragment length polymorphism). A total of 43 sequence-based PCR markers were obtained and polymorphic information content(PIC) was analyzed to measure the informativeness of each marker in watermelon varieties. The average PIC value of SCAR markers was 0.41, which was similar to that of SSR markers. Genetic diversity was also estimated by using these markers to assess the phylogenetic relationships among commercial varieties of watermelon. These markers differentiated 26 Korean watermelon varieties into two major phylogenetic groups, but this grouping was not significantly correlated with their morphological and physiological characteristics. The mean genetic similarity was 66% within the complete set of 26 commercial varieties. In addition, these sequence-based PCR markers were reliable and useful to identify cultivars and genotypes of watermelon.

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Lung Microbiome Analysis in Steroid-Naïve Asthma Patients by Using Whole Sputum

  • Jung, Jae-Woo;Choi, Jae-Chol;Shin, Jong-Wook;Kim, Jae-Yeol;Park, In-Won;Choi, Byoung Whui;Park, Heung-Woo;Cho, Sang-Heon;Kim, Kijeong;Kang, Hye-Ryun
    • Tuberculosis and Respiratory Diseases
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    • v.79 no.3
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    • pp.165-178
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    • 2016
  • Background: Although recent metagenomic approaches have characterized the distinguished microbial compositions in airways of asthmatics, these results did not reach a consensus due to the small sample size, non-standardization of specimens and medication status. We conducted a metagenomics approach by using terminal restriction fragment length polymorphism (T-RFLP) analysis of the induced whole sputum representing both the cellular and fluid phases in a relative large number of steroid $na{\ddot{i}}ve$ asthmatics. Methods: Induced whole sputum samples obtained from 36 healthy subjects and 89 steroid-$na{\ddot{i}}ve$ asthma patients were analyzed through T-RFLP analysis. Results: In contrast to previous reports about microbiota in the asthmatic airways, the diversity of microbial composition was not significantly different between the controls and asthma patients (p=0.937). In an analysis of similarities, the global R-value showed a statistically significant difference but a very low separation (0.148, p=0.002). The dissimilarity in the bacterial communities between groups was 28.74%, and operational taxonomic units (OTUs) contributing to this difference were as follows: OTU 789 (Lachnospiraceae), 517 (Comamonadaceae, Acetobacteraceae, and Chloroplast), 633 (Prevotella), 645 (Actinobacteria and Propionibacterium acnes), 607 (Lactobacillus buchneri, Lactobacillus otakiensis, Lactobacillus sunkii, and Rhodobacteraceae), and 661 (Acinetobacter, Pseudomonas, and Leptotrichiaceae), and they were significantly more prevalent in the sputum of asthma patients than in the sputum of the controls. Conclusion: Before starting anti-asthmatic treatment, the microbiota in the whole sputum of patients with asthma showed a marginal difference from the microbiota in the whole sputum of the controls.

Species Identification of Nontuberculous Mycobacteria (NTM) by PCR-Restriction Fragment Length Polymorphism (PRA) of the rpoB Gene from Three Hospitals of Busan-Kyeongnam Area

  • Choi, Sung-Ran;Kang, Min-Jung;Park, Gyu-Hwan;Kim, Da-Hye;Jeong, Da-Woon;Seo, Eun-Hye;Lee, Hyang-Min;Park, Hyun-Kyung;Jeong, Jin-Yee;Lee, Jung-Min;Jeong, Soo-Young;Lee, Jun-Young;Cho, Eun-Jin;Jekal, Suk;Kim, Chung-Hwan
    • Korean Journal of Clinical Laboratory Science
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    • v.45 no.2
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    • pp.48-53
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    • 2013
  • Recently, the isolation rate of nontuberculous mycobacteria (NTM) in clinical laboratories and the incidence of NTM infections are on the increase in Korea, but there have been only a few studies that reveal the general aspect of NTM isolation or species distribution. Therefore, this study was performed to examine the species identification by PCR-restriction fragment length polymorphism analysis (PRA, PCR-RFLP), and the clinical significance of mycobacterial cultures. PRA was used during the novel region of the rpoB gene and was developed for rapid and precise identification of mycobacteria to the species level. From January 2012 to April 2012, we examined pre-identified nontuberculous mycobacteria (60 species in 3 hospital of Busan-Kyeongnam area). We confirmed 4 (6.6%) Mycobacterium tuberculosis (MTB) and 56 (93.4%) NTM from 60 pre-identified NTM species by multiplex PCR (MolecuTech $MTB-ID^R$ V3, YD Diagnostics, Korea) and PRA (Myco-ID, YD Diagnostics, Korea). The distribution of 56 NTM species were M. intracellulare type I 15 (26.7%), M. avium 14 (25%), M. abscessus 11 (19.5%), M. kansasii type I 3 (5.4%), M. pulveris 2 (3.6%), M. intracellulare type, M. chelonae, M. kansasii type V, M. gallinarum, M. wolinskyi. Respectively, 1 (1.8%) and 6 (10.7%) species were not identified.

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