• Title/Summary/Keyword: DNA Fingerprints

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Analysis of Plasmid pJP4 Horizontal Transfer and Its Impact on Bacterial Community Structure in Natural Soil

  • KIM TAE SUNG;KIM MI SOON;JUNG MEE KUM;JOE MIN JEONG;AHN JAE HYUNG;OH KYOUNG HEE;LEE MIN HYO;KIM MIN KYUN;KA JONG OK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.376-383
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    • 2005
  • Alcaligenes sp. JMP228 carrying 2,4­dichlorophenoxyacetic acid (2,4-D) degradative plasmid pJP4 was inoculated into natural soil, and transfer of the plasmid pJP4 to indigenous soil bacteria was investigated with and without 2,4-D amendment. Plasmid pJP4 transfer was enhanced in the soils treated with 2,4-D, compared to the soils not amended with 2,4-D. Several different transconjugants were isolated from the soils treated with 2,4-D, while no indigenous transconjugants were obtained from the unamended soils. Inoculation of the soils with both the donor Alcaligenes sp. JMP228/pJP4 and a recipient Burkholderia cepacia DBO 1 produced less diverse transconjugants than the soils inoculated with the donor alone. Repetitive extragenic palindromic-polymerase chain reaction (REP-PCR) analysis of the transconjugants exhibited seven distinct genomic DNA fingerprints. Analysis of 16S rDNA sequences indicated that the transconjugants were related to members of the genera Burkholderia and Pandoraea. Denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA genes revealed that inoculation of the donor caused clear changes in the bacterial community structure of the 2,4-D­amended soils. The new 16S rRNA gene bands in the DGGE profile corresponded with the 16S rRNA genes of 2,4-D­degrading transconjugants isolated from the soil. The results indicate that introduction of the 2,4-D degradative plasmid as Alcaligenes sp. JMP228/pJP4 has a substantial impact on the bacterial community structure in the 2,4-D-amended soil.

Genetic Diversity of Paecilomyces japonica and Cordypces militaris Strains by URP-PCR Fingerprinting (URP-PCR핵산지문에 의한 눈꽃동충하초 (Paecilomyces japonica.)와 번데기동충하초(Cordypces militaris) 유전적 다양성분석)

  • Kim, Jong-Kun;Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.39 no.3
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    • pp.180-184
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    • 2011
  • This study was carried out to identify the genetic characteristics among isolates of Paecilomyces spp.and Cordyceps spp. by URP-PCR analysis. Twenty URP (universal rice primer) primers of 20 mer which were designed from repetitive sequence of rice, were used for producing PCR DNA fingerprints of the mushrooms. Of them, 5 URP primers, URP2F, URP2R, URP9F, URP4R, and URP17R amplified genomic DNA of the mushrooms with polymorphic PCR patterns. On isolates of Cordyceps militaris, primers URP1F, URP2R, URP6R and URP17R produced PCR polymorphic bands of 4 types. Isolates of Cordypces sp. that are isolated from different area of Korea were identical to isolate of C. militaris, while other species of Cordypces were different to the PCR profiles. However, the URP primers did not identify the polymorphism of PCR profile on isolates of P. japonica.

Effect of Transgenic Genotype on Transgene Expression in Mud Loach (Misgurnus mizoIepis): I. Copy Number-Dependent Expression in Gynogenetically Derived Homozygous Transgenics

  • Nam Yoon Kwon;Noh Jae Koo;Kim Dong Soo
    • Fisheries and Aquatic Sciences
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    • v.4 no.1
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    • pp.39-46
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    • 2001
  • To examine the effect of copy number-dependent transgenic genotype on the expression of foreign gene, stable hemizygous and homozygous transgenic breeding line was established using artificial parthenogenesis. For this purpose, induced diploid gynogenetic transgenesis was optimized in mud loach (Misgurnus mizolepis) using UV-irradiated cyprinid loach (M. anguillicaudatus) sperm and thermal shocks. Optimum UV range for inactivation of cyprinid loach sperm was between 3,150 to $4,050\;ergs/mm^2$ The UV-irradiated sperm were inseminated into eggs from recessive color strain (yellow) or heterozygous transgenic mud loach containing CAT gene. Cold shock at $2^{\circ}C$ for 60 min, 5 min post fertilization successfully restored the diploidy of eggs inseminated with UV-irradiated sperm. Restoration to diploidy was confirmed by flow cytometry and gynogenetic status was verified by examining maternal exclusive inheritance of multi-locus DNA fingerprints, body color and transgenic marker. Putative isogenic transgenic fish clearly showed homozygous status at trans gene locus based on Southern blot hybridization and progeny testing. Further, such homozygous gynogenetic diploids revealed the increased levels of transgene expression, when compared to those of heterozygous (hemizygous) transgenic fish.

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Molecular fingerprinting of olive flounder pathogenic Streptococcus parauberis strains by random amplified polymorphic DNA analysis

  • Jung, Yong-Uk;Kang, Sang-Hyuck;Jin, Chang-Nam;Kang, Bong-Jo;Heo, Moon-Soo
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.899-903
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    • 2005
  • Two infectious species of Streptococcosis pathogens were detected by multiplex PCR assay. Detection rates of Streptococcus iniae and S. parauberis could reach 44.9% and 55.1% respectively for one year during 2004 to 2005 in Jeju island. These findings showed that S. parauberis strains were important pathogen with streptococcosis of olive flounder in Jeju island. These findings showed that S. parauberis strains were important pathogen with streptococcosis of olive flounder in Jeiu island. In the present study we have investigated the interspecific relationship of all Jeju area of S. parauberis by RAPD analysis. Represent strains divided to four groups by RAPD fingerprints. The important differences observed between the olive flounder isolates suggest that they could constitute a well-differentiated group or a separate clonal line within this bacterial species. Though, serological research of S. parauberis strains in Jeju island not exist yet. These strains doing the serological evolution.

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Lane Detection and Tracking Algorithm for 3D Fluorescence Image Analysis (3D 형광이미지 분석을 위한 레인 검출 및 추적 알고리즘)

  • Lee, Bok Ju;Moon, Hyuck;Choi, Young Kyu
    • Journal of the Semiconductor & Display Technology
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    • v.15 no.1
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    • pp.27-32
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    • 2016
  • A new lane detection algorithm is proposed for the analysis of DNA fingerprints from a polymerase chain reaction (PCR) gel electrophoresis image. Although several research results have been previously reported, it is still challenging to extract lanes precisely from images having abrupt background brightness difference and bent lanes. We propose an edge based algorithm for calculating the average lane width and lane cycle. Our method adopts sub-pixel algorithm for extracting rising-edges and falling edges precisely and estimates the lane width and cycle by using k-means clustering algorithm. To handle the curved lanes, we partition the gel image into small portions, and track the lane centers in each partitioned image. 32 gel images including 534 lanes are used to evaluate the performance of our method. Experimental results show that our method is robust to images having background difference and bent lanes without any preprocessing.

Korea Coast Guard's Human Biological Materials Storage Project for Identifying Bodies Recovered from the Sea: A Model Suggestion (해양변사자 신원확인을 위한 해양경찰의 인체유래물 보관사업 모델제시)

  • Joo, Hyunjung;Choo, Minkyu;Baek, Yoongi;Kim, Namyul;Choi, A-jin;Im, Sonyoung;Lee, Jongnam;Kim, Hyungkyu;Lee, Hanseong
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.24 no.2
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    • pp.171-178
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    • 2018
  • The ocean is a harsh environment and Korea Coast Guard officers often face the threat of death or going missing in the line of duty. In order to promptly identify unknown bodies during mishaps, Korea Coast Guard Research Center has established an identification system using the DNA of its officers. This collected DNA can also be entombed in National Cemeteries when the remains are not recovered. The storage, disposal and quality control of the materials are overseen by the Governing Committee according to the Enforcement Rules for the Human Biological Materials Storage Project for the Identification of Officers Killed or Gone Missing in Action. Approximately 700 bodies are found per year along the Korean coast. An alternative method should be put in place for when identification through fingerprints and teeth are not applicable due to severe decomposition or partial recovery. We believe it would greatly contribute to helping identify the unknown bodies if the storage project could expand its data to include marine industry workers, relevant researchers and those involved in marine leisure activities.

Comparison of Biological Characteristics on the Organic Waste-treated Lysimeter Soil by RFLP, PLFA, and CLSU (RFLP, PLFA, CLSU를 이용한 폐기물연용토양의 토양미생물 특성 평가 비교)

  • Jang, Kab-Yeul;Weon, Hang-Yeon;Lee, Kang-Hyo;Kwon, Sun-Ik;Kong, Won-sik;Suh, Jang-sun;Sung, Jae-Mo
    • Korean Journal of Soil Science and Fertilizer
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    • v.41 no.6
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    • pp.415-418
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    • 2008
  • The application of sludge wastes into agricultural fields has been increasing annually in Korea. In particular, sewage sludge application has been widely accepted in decades. Sewage sludge application aid in the recycling of essential nutrients and act as a source of organic matter improving the structure and water-holding properties of the soil. The efficient use of sludge wastes, however, requires an individual assessment of waste products. This study assessed the biological characteristics of organic waste-treated lysimeter soils and develop its indicator to assess the soil health of organic waste-treated lysimeter soils. Several analytical techniques more recently developed such as restriction fragment length polymorphism (RFLP), phospholipid fatty acid (PLFA), and community level substrate utilization (CLSU) fingerprints allow for detailed analyses of soil microbial communities. PLFA and RFLP was, therefore, used in the study to characterize the microbial communities in soil without the need to isolate individual fungi and bacteria. PLFA, RFLP and CLSU have been utilized to assess microbial characteristics of the lysimeter soils with four different sludge wastes for eight consecutive years. Each of these methods was analyzed for a different aspect of soil microbial characteristics. The study would disclose those methods yielded highly reproductive results for each soil and allow distinguishing the soils based on the structures of specific geneand PLFA-pools more than CLSU fingerprints. PLFA methods, especially, revealed the same relative similarities of the treated soils based on cluster analysis of the biological characteristics. Pig manure compost-treated soil, however, was only the same relative resemblance among the three methods. These results indicated that PLFA easily assessed the biological soil characterization.

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.

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.

Bacterial Diversity of the South Pacific Sponge, Dactylospongia metachromia Based on DGGE Fingerprinting (DGGE에 의한 남태평양 해면 Dactylospongia metachromia의 공생세균 다양성)

  • Jeong, In-Hye;Park, Jin-Sook
    • Korean Journal of Microbiology
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    • v.49 no.4
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    • pp.377-382
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    • 2013
  • The bacterial community structures of the marine sponge, Dactylospongia metachromia, collected from Chuuk of Micronesia on February 2012, were analyzed by denaturing gradient gel electrophoresis (DGGE). The DGGE fingerprints of two individuals of D. metachromia, CH607 and CH840 showed the same band patterns. The sequences derived from DGGE bands revealed 93~100% similarities with known bacterial species in the public database and high similarity with uncultured bacterial clones. The bacterial community structures of both D. metachromia sponges (CH607, CH840) were composed of 6 phyla, 8 classes: Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Acidobacteria, Actinobacteria, Chloroflexi, Cyanobacteria, Spirochaetes. DGGE fingerprint - based phylogenetic analysis revealed that the bacterial community profiles were identical in two individuals of the same sponge species collected from the same geographical location.