• Title/Summary/Keyword: DNA fingerprinting

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Analysis of DNA fingerprints of Mycobacterium Tuberculosis Isolates from Patients Registered at Health Center in Gyeonggi Province in 2004 (2004년도 경기도 보건소 결핵환자로 부터 분리된 결핵균 DNA 지문분석)

  • Park, Young Kil;Kang, Hee Yeun;Lim, Jang Geun;Ha, Jong Sik;Jo, Jung Ok;Choi, Hang Soon;Lee, Ka Chel;Choi, Young Hwa;Sheen, Seung Soo;Jeon, Gi-Hong;Bai, Gil Han
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.3
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    • pp.290-296
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    • 2006
  • Background : IS6110 DNA fingerprint is a very useful tool for investigating the transmission of tuberculosis. The aim of this study was to identify the epidemiological situations within a given area (one province). Methods : The 681 Mycbobacterium tuberculosis isolates from patients, who were registered at health centers in Gyeonggi Province from May to December in 2004, were subjected to IS6110 DNA fingerprinting. Patients belonging to clusters were interviewed by health-workers to determine their previous contacts or household TB history. Results : The number of IS6110 copies of the 681 isolates showed diverse fingerprint patterns from 0 to 21 of which the most prevalent copy number was 10 from 120 isolates (17.6%). Thirty-three isolates (4.8%) belonged to the K strain, and 128 isolates (18.8%) belonged to the K family. There were 180 (26.4%) isolates belonged belonging to fifty clusters, of which two clusters were within household transmission. Forty-three (23.9%) out of 180 patients resided in an area under the same health center control. The rate of clusters in those aged 60-70 was higher than in any other age group ( 95% CI of RR : 1.072 ~ 1.988). Conclusion : This is the first report of an epidemiological survey based on a whole province using a DNA fingerprinting technique for M. tuberculosis. These results will be helpful in developing a program or policies to prevent the transmission of TB.

Cellular responses and proteomic analysis of hemolytic Bacillus cereus MH-2 exposed to epigallocatechin gallate (EGCG) (Epigallocatechin Gallate (EGCG)에 노출된 용혈성 Bacillus cereus MH-2의 세포 반응 및 프로테옴 분석)

  • Kim, Dong-Min;Park, Sang-Kook;Oh, Kye-Heon
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.260-268
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    • 2016
  • The aim of this work was to investigate the cellular responses and proteomic analysis of Bacillus cereus MH-2 exposed to EGCG. Strain MH-2 was isolated from commercial Ssamjang and has the hemolytic activity. Survival of the MH-2 strain with time in the presence of different concentrations of EGCG under sublethal conditions was monitored. The amount of alginate from MH-2 strain decreased depending on the increasing concentrations of EGCG and increased depending on the exposure time at any particular EGCG concentration. Analysis of SDS-PAGE and Western blot using anti-DnaK and anti-GroEL revealed that two stress shock proteins, 70 kDa DnaK and 60 kDa GroEL were found to decrease in proportion to the EGCG concentration in exponentially growing cultures. Scanning electron microscopic analysis demonstrated the presence of protrusions and fused rod forms on the cells treated with EGCG. 2-DE of soluble protein fractions from MH-2 cultures showed 20 protein spots changed by EGCG exposure. These proteins involved in enterotoxins (hemolysin BL lytic component L1 and hemolysin BL-binding protein), chaperons (DnaK and GroEL), cell defense (peptidase M4 family proteins), and various biosynthesis and energy metabolism were identified by peptide mass fingerprinting using MALDI-TOF. These results provide clues for understanding the mechanism of EGCG-induced stress and cytotoxicity on B. cereus MH-2.

Isolation of Cryptococcus neoformans var. grubii (serotype A) from Pigeon Droppings in Seoul, Korea

  • Chee, Hee-Youn;Lee, Kyung-Bok
    • Journal of Microbiology
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    • v.43 no.5
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    • pp.469-472
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    • 2005
  • Seventy-two pigeon dropping samples were collected from 26 different localities in Seoul and investigated for the occurrence of Cryptococcus neoformans. Seventeen samples from 8 different localities were found to be positive for C. neoformans. All isolates were obtained from withered pigeon droppings. Identification and serotyping of the isolates were determined by means of serological testing and DNA fingerprinting. All isolates belonged to C. neoformans var. grubbi (serotype A).

DNA Profiles of Trichoderma spp. in Korea

  • Park, Dong-Suk;Kang, Hee-Wan;Park, Young-Jin;Lee, Mi-Hee;Lee, Byoung-Moo;Hahn, Jang-Ho;Go, Seung-Joo
    • Mycobiology
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    • v.32 no.1
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    • pp.24-34
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    • 2004
  • Molecular approaches, internal transcribed spacer(ITS) sequences of ribosomal DNA, and Universal Rice Primer Polymerase Chain Reaction(URP-PCR) were used to investigate the genetic diversity, taxonomic complexity, and relationships of Trichoderma species in mushroom farms. Forty-one isolates of 13 Trichoderma spp. were used in this study and clustered into eight groups. The DNA fingerprint patterns and ITS1 region sequence alignment data showed similar results, but not in some species, such as T. virens, T. atroviride, T. harzianum, and T. aureoviride. Results of this study have proven that the morphology-based taxonomic system has some limitations in terms of classification. The data obtained in this study would be a good index for classifying indistinguishable Trichoderma strains.

Oropharyngeal Carriage and Antimicrobial Resistance of S. pneumoniae in Children of Seoul (서울 지역 소아의 구인강에서 폐구균 보균율과 항균제 내성)

  • Kim, Young Kee;Lee, Chang Kyu
    • Pediatric Infection and Vaccine
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    • v.4 no.2
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    • pp.218-224
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    • 1997
  • Purpose: The antimicrobial resistance of S. pneumoniae has encountered with increasing frequency from around the world. In our country, penicillin resistant strains of S. penumococci are rapidly increasing. It has been known that colonized pneumococci in upper respiratory tract cause sinisitis, otitis media, meningitis and pneumonia. We tried to reveal the colonization rate of pneumonocci in upper respiratory tract, their antimicrobial resistance and DNA fingerprinting pattern in normal children. Methods: We got specimens from 117 children of day-care center in Seoul through oropharyngeal swab. After incubation on BAP, optochin test and slide latex agglutination test were used for identification. Antimicobial susceptibility test to penicillin, vancomycin, erythromycin and TMP-SMZ was done with disk diffusion method. Penicillin MIC was gotten through the broth microdilution method. Genotyping of 45 pneumococci was done by rep-PCR using REP1R-Dt and REP2-Dt primer. Results: The carriage rate of pneumococci in the day-care center children was 38%(45/117). The resistance of penicillin, erhthromycin, TMP/SMZ, vancomycin by the disk diffusion method are 89%, 91%, 64% and 0%, respectively. 64% of the isolates showed multiple resistance. 7 types of DNA fingerprinting were gotten and 78% of isolates belonged to three types. Conclusion: We found that the antimicrobial resistance of children attending the day-care center in Seoul was much higher than expected. We assumed that this might be due to their easy and frequent exposure to antimicrobial agents and crowded day-care center environment.

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DNA fingerprinting analysis for soybean (Glycine max) varieties in Korea using a core set of microsatellite marker (핵심 Microsatellite 마커를 이용한 한국 콩 품종에 대한 Fingerprinting 분석)

  • Kwon, Yong-Sham
    • Journal of Plant Biotechnology
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    • v.43 no.4
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    • pp.457-465
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    • 2016
  • Microsatellites are one of the most suitable markers for identification of variety, as they have the capability to discriminate between narrow genetic variations. The polymorphism level between 120 microsatellite primer pairs and 148 soybean varieties was investigated through the fluorescence based automatic detection system. A set of 16 primer pairs showed highly reproducible polymorphism in these varieties. A total of 204 alleles were detected using the 16 microsatellite markers. The number of alleles per locus ranged from 6 to 28, with an average of 12.75 alleles per locus. The average polymorphism information content (PIC) was 0.86, ranging from 0.75 to 0.95. The unweighted pair group method using the arithmetic averages (UPGMA) cluster analysis for 148 varieties were divided into five distinctive groups, reflecting the varietal types and pedigree information. All the varieties were perfectly discriminated by marker genotypes. These markers may be useful to complement a morphological assessment of candidate varieties in the DUS (distinctness, uniformity and stability) test, intervening of seed disputes relating to variety authentication, and testing of genetic purity in soybean varieties.

Structure and Diversity of Arsenic-Resistant Bacteria in an Old Tin Mine Area of Thailand

  • Jareonmit, Pechrada;Sajjaphan, Kannika;Sadowsky, Michael J.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.169-178
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    • 2010
  • The microbial community structure in Thailand soils contaminated with low and high levels of arsenic was determined by denaturing gradient gel electrophoresis. Band pattern analysis indicated that the bacterial community was not significantly different in the two soils. Phylogenetic analysis obtained by excising and sequencing six bands indicated that the soils were dominated by Arthrobacter koreensis and $\beta$-Proteobacteria. Two hundred and sixty-two bacterial isolates were obtained from arsenic-contaminated soils. The majority of the As-resistant isolates were Gramnegative bacteria. MIC studies indicated that all of the tested bacteria had greater resistance to arsenate than arsenite. Some strains were capable of growing in medium containing up to 1,500 mg/l arsenite and arsenate. Correlations analysis of resistance patterns of arsenite resistance indicated that the isolated bacteria could be categorized into 13 groups, with a maximum similarity value of 100%. All strains were also evaluated for resistance to eight antibiotics. The antibiotic resistance patterns divided the strains into 100 unique groups, indicating that the strains were very diverse. Isolates from each antibiotic resistance group were characterized in more detail by using the repetitive extragenic palindromic-PCR (rep-PCR) DNA fingerprinting technique with ERIC primers. The PCR products were analyzed by agarose gel electrophoresis. The genetic relatedness of 100 bacterial fingerprints, determined by using the Pearson product-moment similarity coefficient, showed that the isolates could be divided into four clusters, with similarity values ranging from 5-99%. Although many isolates were genetically diverse, others were clonal in nature. Additionally, the arsenic-resistant isolates were examined for the presence of arsenic resistance (ars) genes by using PCR, and 30% of the isolates were found to carry an arsenate reductase encoded by the arsC gene.

Genetic Diversity of Korean Rice Breeding Parents as Measured by DNA Fingerprinting with Simple Sequence Repeat (SSR) Markers

  • Song, Moon-Tae;Lee, Jeom-Ho;Lee, Sang-Bok;Cho, Youn-Sang;Ku, Ja-hwan;Seo, Kyoung-In;Choi, Seong-ho;Hwang, Heung-Goo
    • Plant Resources
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    • v.6 no.1
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    • pp.16-26
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    • 2003
  • Molecular markers are useful tools for evaluating genetic diversity and determining cultivar identity. Present study was conducted to evaluate the genetic diversity within a diverse collection of rice accessions used for Korean breeding programs. Two hundred eighty-seven rice cultivars, composed of temperate japonica, tropical japonica, indica, and Tongil-type of Korean crossing parents were evaluated by means of 15 simple sequence repeat (SSR) markers. A total of 99 alleles were detected, and the number of alleles per marker ranged from 4 to 11, with an average of 6.6 per locus. Polymorphism information content (PIC) for each of the SSR markers ranged from 0.2924 to 0.8102 with an average of 0.5785. These results, with the result that use of only 15 SSR markers made all rice cultivars examined could be uniquely distinguished, imply the efficiency of SSR markers for analysis of genetic diversity in rice. Cluster analysis was performed on similar coefficient matrics calculated from SSR markers to generate a dendogram in which two major groups corresponding to japonica (Group I) and indica and Tongil type rice (group II) with additional subclasses within both major groups. The narrowness of the Korean breeding germplasm was revealed by the fact that most of the Korean-bred and Japan-bred temperate japonica cultivars were concentrated into only 2 of the sub-group I-1 (143 cultivars) and I-2 (58 cultivars) among six sub-groups in major group of japonica. This is because of the japonica accessions used in this study was a very closely related ones because of frequent sharing of the crossing parents with similar genetic background with synergy effect of the inherited genetic difference between indica and japonica. A rice breeding strategy with the use of molecular markers was discussed for overcoming of genetic vulnerability owing to this genetic narrowness.

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