• Title/Summary/Keyword: Restriction Fragment Length

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Characterization of Cucumber mosaic virus Isolated from Trifolium repens in Korea (국내 토끼풀에서 분리한 Cucumber mosaic virus의 특성)

  • Park, Tae Seon;Choi, Gug Seoun;Hong, Jin Sung
    • Research in Plant Disease
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    • v.22 no.1
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    • pp.55-58
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    • 2016
  • A Cucumber mosaic virus (named CMV-Tr1) isolated from the white clover (Trifolium repens) showing mosaic and malformation that found in a pepper field. Cucumber mosaic virus was identified through confirmation with PT-PCR, PCR-restriction fragment length polymorphism, and sequence analysis of coat protein (CP) gene. CMV-Tr1 mosaic symptom on the upper leaves of five tobacco species including Nicotiana benthamiana, Cucumis sativus, Physalis angulata, and Solanum lycopersicon. In Chenopodium quinoa and Vigna unguiculata the isolate showed local lesions in inoculated leaves. CMV-Tr1 compared with CMV-As in the sequence identity of CP gene. CMV-Tr1 showed 98.9% and 99.5% homologies at nucleotide and amino acid levels, respectively. Phylogenetic analysis of the CP gene indicated that CMV-Tr1 belongs to the CMV subgroup IB base on the CP. To our knowledge, this is the first report of CMV in T. repens in Korea.

Association of GRIA1 polymorphisms with ovarian response to human menopausal gonadotropin in Iranian women

  • Golestanpour, Hossein;Javadi, Gholamreza;Sheikhha, Mohammad Hasan
    • Clinical and Experimental Reproductive Medicine
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    • v.47 no.3
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    • pp.207-212
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    • 2020
  • Objective: Glutamate ionotropic receptor AMPA type subunit 1 (GRIA1) is a subunit of a ligand-gated ion channel that regulates the secretion of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) by controlling the release of gonadotropin-releasing hormone. Few studies have investigated the association between the GRIA1 gene and human infertility. This study evaluated the association of the GRIA1 rs548294 C > T and rs2195450 G > A polymorphisms with the ovarian response to human menopausal gonadotropin (HMG) in Iranian women. Methods: One hundred women with histories of at least 1 year of infertility were included. On the second day of menstruation, patients were injected with HMG; on the third day, blood samples were collected. After hormonal analysis, the GRIA1 rs548294 C > T and rs2195450 G > A genotypes of samples were identified via the restriction fragment length polymorphism method, and on day 9, the number of follicles was assessed via ultrasound. Results: For the GRIA1 rs548294 C > T and rs2195450 G > A single nucleotide polymorphisms, the subjects with CT and GG genotypes, respectively, displayed the highest mean FSH level, LH level, and number of follicles on day 9 of the menstrual cycle (p< 0.05). Significant positive correlations were observed between LH and FSH (p< 0.01), LH and follicle count (p< 0.01), FSH and age (p< 0.05), follicle count and age (p= 0.048), and FSH and follicle count (p< 0.01). Conclusion: This study showed a significant relationship between GRIA1 polymorphisms and ovarian response to the induction of ovulation. Therefore, determining patients' GRIA1 genotype may be useful for improving treatment and prescribing suitable doses of ovulation-stimulating drugs.

Lack of Association of a Common Polymorphism in the 3'-UTR of Interleukin 8 with Non Small Cell Lung Cancer in Kashmir

  • Bhat, Imtiyaz Ahmad;Pandith, Arshid A.;Bhat, Bashir A.;Naykoo, Niyaz A.;Qasim, Iqbal;Rasool, Roohi;Aziz, Sheik Aejaz;Shah, Zafar Amin
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.7
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    • pp.4403-4408
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    • 2013
  • Background: Chronic inflammation is considered as an important factor in the pathogenesis of lung cancer. The presence of inflammatory cells and higher levels of pro-inflammatory cytokines in the tumor microenvironment and their surrounding tissues is gaining much importance in research. Materials and Methods: One hundred ninety NSCLC cases and 200 age, smoking and sex matched controls were evaluated for association of IL-8 -251 (rs4073) and IL-8 -845 (rs2227532) in our population. Restriction fragment length polymorphism (RFLP) was used followed by direct sequencing for the detection of SNPs. Results: The IL-8 -845 polymorphism was not found in our population. No significant association was observed between the IL-8 -251 AT genotypes and IL-8 -25 AA genotypes and NSCLC (p=0.05) in our population. The IL-8 -251 A allele was also non-significant (p=0.05) in NSCLC patients. Conclusions: In conclusion, this report reveals lack of association between IL-8 - 251 A/T polymorphism and NSCLC in our Kashmir Valley population.

Association of Polymorphisms in Epidermal Growth Factor, Prostaglandin-endoperoxide Synthase 2 and Prolactin Receptor Genes with Semen Quality in Duroc Boars

  • Huang, S.Y.;Song, H.L.;Lin, E.-C.;Lee, W.C.;Chiang, J.C.;Tsou, H.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.6
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    • pp.793-798
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    • 2006
  • The quality characteristics of semen are important indicators of the fertility of a boar. Development of genetic markers for the semen quality in boars will be beneficial to the improvement of porcine fertility. We investigated the relationship between the polymorphisms of epidermal growth factor (EGF), prostaglandin-endoperoxide synthase 2 (PTGS2) and prolactin receptor (PRLR) genes, and semen quality traits in boars. The genomic DNA of 233 boars (157 Duroc and 86 Landrace) from a central testing station was subjected to genotyping for surveying gene frequency. The EGF, PTGS2 and PRLR genotypes were determined using the restriction fragment length polymorphism method. Thirty-seven normal, mature Duroc boars from an AI center were also genotyped and their semen quality traits were collected. The effect of genotype on semen quality traits was analyzed by the least-squares means method using data corrected for season. The frequencies of the AA genotype of EGF, PTGS2 and PRLR in Duroc boars were 0.14, 0.01 and 0.66, respectively. In Landrace, the frequencies of the AA genotype were 0.03, 0.09 and 0.62, respectively. Boars with the BB genotype in EGF, with the AB genotype in PTGS2 and with the AA genotype in PRLR had significantly better semen quality with a higher percentage of normal sperm and a lower percentage of immature sperm than those with other genotypes. These findings imply that polymorphisms of EGF, PTGS2 and PRLR genes might be used as markers for improving the semen quality of boars.

AN EXPERIMENTAL STUDY ON THE CHEMOPREVENTIVE EFFECT OF CHLOROPHYLLIN IN HAMSTER CHEEK POUCH TUMOR INDUCED BY 7, 12-DIMETHYLBENZ[A]ANTHRACENE (7, 12-dimethylbenz[a]anthracene(DMBA)로 유발된 햄스터 협낭암에서 chlorophylln의 암예방효과에 관한 실험적 연구)

  • Yoon, Kyu-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.26 no.2
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    • pp.137-145
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    • 2000
  • Carcinogenesis is a multi-stage process that generally consists of at least three steps; initiation, promotion, and progression. If one of these carcinogenic steps were suppressed or delayed, the cancer could be prevented. Cancer chemoprevention is defined to be inhibition or reversal of the carcinogenic process by the specific chemical agents and is a novel approach to cancer management alternative to conventional chemotherapy. Chlorophylln(CHL), a water-soluble derivative of chlorophyll, containing sodium and copper, has been known to be strong antimutagen in several test systems, but its mechanism of antimutagenic action is unknown. In the present experiment, the possibility of CHL as chemopreventive drugs on 7,12-dimethylbenz[a]anthracene(DMBA)-induced hamster buccal pouch carcinogenesis was investigated by mutagenicity test, carcinogenicity test, and frequency or spectrum of H-ras mutations in the both of DMBA-induced and chlorophylln-pretreated-DMBA induced tumor by polymerase chain reaction and non-isotopic restriction fragment length polymorphism. The treatment of CHL reduced the yields and multiplicity of the 0.5% DMBA-induced tumor, 86% to 62.5% and $3.7{\pm}0.6$ to $1.4{\pm}0.3$, respectively. The occurrence of histidine revertant by $20{\mu}mole$ DMBA was inhibited 25.6 to 81.7% by 1 to $5{\mu}M$ CHL in a dose-dependent manner. The mutation rates of H-ras gene in DMBA-induced and CHL-pretreated-DMBA induced tumor were 96%, 94% of which the most mutations were in codon 12/13. These results suggest that CHL inhibits the carcinogenic action of DMBA by the formation of complex between CHL and DMBA or the inhibition of the activation of DMBA in vivo. But CHL did not affect the mutation rates or its spectrum in already formed tumor.

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Community Analysis of Nitrite-Oxidizing Bacteria in Lab-Scale Wastewater Treatment System (폐수처리장치에서의 아질산염 산화 세균 군집 분석)

  • Jeong, Soon-Jae;Lee, Sang-Ill;Lee, Dong-Hun
    • Korean Journal of Microbiology
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    • v.44 no.1
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    • pp.29-36
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    • 2008
  • Nitrogen is one of the major pollutants that should be removed by wastewater treatment systems. Biological nitrogen removal (BNR) is a key technology in advanced wastewater treatment systems operated by bacterial populations. Nitrification is the first step of microbiological processes in BNR system. Ammonia is oxidized to nitrite by ammonia-oxidizing bacteria (AOB) and then nitrite is subsequently oxidized to nitrate by nitrite-oxidizing bacteria (NOB). The diversity of NOB in nitrification reactors of 3 BNR systems, Edited biological aerated filter system, Nutrient removal laboratory system, and the Rumination type sequencing batch reactor system, was investigated by terminal restriction fragment length polymorphism (T-RFLP) analysis of 16S rRNA genes. Cluster analysis of T-RF profiles showed that communities of Nitrobacter group in each system were different depending upon the process of systems. However, the clusters of Nitrospira group were divided by the habitat of aqueous and solid samples.

Bacterial Community Monitoring of Commercial Kimchi Produced in Korea and China with Evidence of Bacilli Spore Formation during Fermentation (한국산 및 중국산 김치의 Bacteria 군집 분석 및 발효과정 중 Bacilli 포자 형성 규명)

  • An, Doohyun;Kim, Hye-Rim;Jeong, Do-Won;Caldwell, Jane M.;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.42 no.2
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    • pp.121-130
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    • 2014
  • Terminal Restriction Fragment Length Polymorphism (T-RFLP) analysis was adopted to explore rapid differentiation in the diversity and dynamics of bacteria in kimchi made in Korea and China for future application in kimchi origin discrimination. T-RFLP analysis supported the reproducible and rapid detection of major lactic acid bacteria known to be involved in kimchi fermentation. The taxonomic resolution level of this T-RFLP analysis was between the species and genus level, but was not specific enough for the detection of a bacterium found only in one origin, either Korea or China. The bacterial community structure successions in kimchi samples from Korea and China analyzed by T-RFLP analysis occurred with a similar pattern. Bacillus spp. which were not detected in the early microbial studies of kimchi were constantly detected until the late fermentation stage of kimchi in our T-RFLP analysis and their existence was proved by culture-based identification. Additionally, sporulation of Bacillus spp. during kimchi fermentation was discovered.

T-RFLP Analysis of Microbial Community Structure in Leachate from Landfill Sites (폐기물매립장 침출수내 미생물군집 구조 해석을 위한 T-RFLP의 활용)

  • Yu, Jae-Cheul;Ishigaki, Tomonori;Kamagata, Yoichi;Lee, Tae-Ho
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.4
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    • pp.369-378
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    • 2010
  • Microorganisms are key-role player for stabilization of landfill sites. In order to evaluate the availability of T-RFLP(Terminal Restriction Fragment Length Polymorphism) for monitoring microbial community variations during stabilization of landfill sites, the phylogenic diversity of microbial community in the leachate from 4 different full-scale landfills was characterized by T-RFLP based on bacterial 16S rDNA. Main population of microbial community analyzed by T-RFLP was significantly similar with that of microbial community analyzed by clone library analysis. The results of T-RFLP analysis for main population of microbial community in the leachate from landfills with different landfill structures, waste types and landfill ages showed apparently different microbial diversity and structures. Therefore, long-term monitoring of microbial community in leachate from landfill sites by using T-RFLP is expected to be available for evaluation of landfill stability.

Cyanobacterial Diversity Analysis Using cpcBA-Intergenic Spacer Region (cpcBA-Intergenic Spacer Region을 이용한 Cyanobacteria의 다양성 분석)

  • Choi Gang-Guk;Park Yong-Ha;Ahn Chi-Yong;Bae Myoung-Sook;Oh Hee-Mock
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.287-292
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    • 2005
  • The cyanobacterial diversity was analyzed by restriction fragment length polymorphism (RFLP) of PCR-amplified rpcBA-Intergenic Spacer (IGS) genes and cpcBA-IGS gene sequencing with a sample collected at Chuso-ri in Daechung Reservoir on March 15, 2005, The Shannon-Weiner diversity index was 0.65, indicating that the cyanobacterial community structure was simple. PCR-RFLP profiles obtained were Phormidium spp. (58 clones), Anabaena spp. (14 clones), Microcystis spp. (4 clones), Spirulina sp. (1 clone) and uncultured cyanobacteria (2 clones). The PCR-RFLP of cpcBA-IGS revealed that Phormidium spp. and Anabaena spp. dominated in the invested sample. As a consequence, it seems that the analysis of functional genes such as cpcBA-IGS can be used for the species identification and community analysis of cyanobacteria.

Molecular Detection and Phylogenetic Analysis of Anaplasma phagocytophilum in Horses in Korea

  • Seo, Min-Goo;Ouh, In-Ohk;Choi, Eunsang;Kwon, Oh-Deog;Kwak, Dongmi
    • Parasites, Hosts and Diseases
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    • v.56 no.6
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    • pp.559-565
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    • 2018
  • The identification and characterization of pathogenic and zoonotic tick-borne diseases like granulocytic anaplasmosis are essential for developing effective control programs. The differential diagnosis of pathogenic Anaplasma phagocytophilum and non-pathogenic A. phagocytophilum-like Anaplasma spp. is important for implementing effective treatment from control programs. The objective of the present study was to investigate the prevalence of Anaplasma spp. in horses in Korea by nucleotide sequencing and restriction enzyme fragment length polymorphism assay. Of the 627 horses included in the study, only 1 (0.2%) was infected with A. phagocytophilum. Co-infection with A. phagocytophilumlike Anaplasma spp. was not detected in the study. The 16S rRNA sequence of A. phagocytophilum was similar (99.5-100%) to A. phagocytophilum 16S rRNA isolated from horses in other countries. PCR adapted to amplify A. phagocytophilum groEL and msp2 genes failed to generate amplicons, suggesting genetic diversity in these genes. This study is the first molecular detection of A. phagocytophilum in horses in Korea. Human granulocytic anaplasmosis and animal infection of A. phagocytophilum have been reported in Korea recently. Because of vector tick distribution, global warming, and the increase of the horse industry, horses should be considered as a potential reservoir for A. phagocytophilum, and cross infectivity should be evaluated even though a low prevalence of infection was detected in this study. Furthermore, continuous surveillance and effective control measures for A. phagocytophilum should be established to prevent disease distribution and possible transmission to humans.