• Title/Summary/Keyword: PCR 동정

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Isolation and Diversity of Wild Yeasts from the Waters and Bank Soils of Daejeoncheon, Gapcheon, and Yudeungcheon in Daejeon Metropolitan City, Korea (대전광역시 대전천과 갑천 및 유등천의 물과 주변 토양들로부터 야생효모들의 분리 및 분포 특성)

  • Han, Sang-Min;Lee, Sang-Yeop;Kim, Ha-Kun;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.45 no.4
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    • pp.259-269
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    • 2017
  • In order to investigate the diversity of yeasts from major rivers (the Daejeoncheon, Gapcheon and Yudeungcheon) located in Daejeon city, we isolated wild yeasts by plating diluents of samples collected during the summer and winter of 2016 onto yeast extract-peptone-dextrose (YPD) medium, then identified them using Basic Local Alignment Search Tool (BLAST) analysis to compare the nucleotide sequences of the PCR amplicons for the D1/D2 domain of 26S rDNA. In total, we isolated 191 yeast strains belonging to 104 species from 148 soil or water samples from the rivers and their junctions. Candida spp. (45 strains) including Candida tropicalis (22 strains) were the most abundantly isolated strains from the Daejeoncheon. Candida spp. (16 strains) including Candida vartiovaarae (8 strains) and Candida spp. (18 strains) such as Candida sake (4 strains) were also the dominant isolates from the Gapcheon and Yudeungcheon, respectively. In conclusion, Candida spp. and Cryptococcus spp. were the most dominant strains, corresponding to 42% and 7% of the 191 yeast strains isolated in this study, respectively.

Analysis of Bacterial Community Structure in Gossi Cave by Denaturing Gradient Gel Electrophoresis (DGGE) (DGGE를 이용한 동굴 생태계 세균 군집 구조 분석)

  • 조홍범;정순오;최용근
    • Korean Journal of Environmental Biology
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    • v.22 no.1
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    • pp.213-219
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    • 2004
  • The bacterial community of water stream, soil and guano in Gossi cave was examined by using PCR amplified the 16S rDNA-denaturing gradient gel electrophoyesis (DGGE). In this study, the genetic diversity and the similarity of bacterial community between open area and non - open area toy cave tour were investigated, and the seasonable variation pattern was compared each other. DGGE is attractive technique, as it sepayate same length dsDNA according to sequence variation typical 16S rDNA genes. The diversity and similarity of bacterial community in cave was analyzed by GC341f and PRUN518r primer sets foy amplification of V3 region of eubacteria 16S rDNA. The specific DGGE band profile of the cave water gives the possibility that the specific bacterial cell can be adapting to the specific cave environment and living in the cave. The DGGE band profiles of all samples with guano were compared and analyzed by image analyzer, in which mutual band profile was compared to be and the band intensity of guano was the highest. From these result, it is thought that the guano was main nutrient source and influenced on the community structure of the cave environment where is nutritionally limited. Pseudomonas sp. NZ060, Pseudomonas pseudoalcaligenes, uncultured Variovorax sp. and soli bacterium NS7 were identified to be on some sample from analysing DNA sequence of some DGGE band.

Genomic Species Identification of Acinetobacter calcoaceticus - Acinetobacter baumannii Complex Strains by Amplified Ribosomal DNA Restriction Analysis (ARDRA) (Amplified Ribosomal DNA Restriction Analysis (ARDRA) 방법을 이용한 국내 분리 Acinetobacter calcoaceticus - Acinetobacter baumannii Complex 균주의 유전자종 동정)

  • Oh, Jae-Young;Cho, Jae-We;Park, Jong-Chun;Lee, Je-Chul
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.1
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    • pp.69-76
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    • 2000
  • Members of the genus Acinetobacter are recognized as newer pathogens of the nosocomial infection with an increasing frequency in recent years. Strains that belonged to A. calcoaceticus A. baumannii complex (genomic species 1, 2, 3, and 13TU) were major groups associated with nosocomial infection. Phenotypic identification was unreliable and laborious method to classify Acinetobacter strains into 19 genomic species. Rapid and reliable identification of clinical isolates is essential to diagnosis and epidemiology of Acinetobacter. We investigated the suitability of amplified ribosomal DNA restriction analysis (ARDRA) to identify genomic species of 131 Acinetobacter isolates. The 16S rRNA genes (ribosomal DNA) were enzymatically amplified and the amplified PCR products were restricted independently with the enzymes, AluI, CfoI, and MboI. Genomic species of Acinetobacter was classified by the combinations of restriction patterns. The analysis was showed that restriction profiles were characteristic for each genomic species. One hundred fourteen isolates were identified as A. baumannii, twelve were identified as genomic species 13TU, and one was identified as genomic species 3. Four isolates were found to be unknown organisms. All of the isolates which were identified to A. baumannii by phenotypic tests were completely discriminated into A. baumannii and genomic species 13TU by ARDRA. This study demonstrates that ARDRA is a rapid and simple techniques for the identification of Acinetobacter species according to the genomic species.

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Multisystemic Aspergillosis with Granulomas in Layer Chickens (산란계에서의 전신 다발성 육아종성 Aspergillosis 발생)

  • Kim, Ji-Ye;Kim, Jong-Man;Mo, In-Pil
    • Korean Journal of Poultry Science
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    • v.38 no.1
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    • pp.45-50
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    • 2011
  • A case of aspergillosis in 39-day-old layer chickens having a history of gradual emaciation and subsequently death with nervous signs such as torticollis and lack of equilibrium was documented. Based on the results from serology and polymerase chain reaction (PCR) test, this flock was not affected with known viral or bacterial diseases. On postmortem examination of the affected birds, multiple white to yellow nodules measuring 1~5 mm in diameter were observed in the lungs, cerebrum, liver and kidney. Microscopically, these nodules were identified as granulomatous lesions characterized by mixed population of multinucleated giant cells and lymphocytes. By periodic acid-schiff staining and nucleotide sequencing analysis, Aspergillus flavus with characteristic septate and branched hyphae were identified in the granuloma of lung and cerebrum. This case was a chronic and multisystemic aspergillosis specialized to central nervous system caused by Aspergillus flavus infection in the layer flocks.

Infection of Clinostomum complanatum (Treamdoda: Digenea) metacercaria in cultured southern top mouthed minnow Pseudorasbora parva (참붕어, Pseudorasbora parva의 인두흡충, Clinostomum complanatum 피낭유충 감염증)

  • Jung, Eun-Bin;Kim, Dong-Wan;Kim, Jin-Do;Park, Sung-Woo
    • Journal of fish pathology
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    • v.25 no.2
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    • pp.85-93
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    • 2012
  • Many metacercariae in the musculature of southern top mouthed minnow (Pseudorasbora parva) cultured at a farm in Chonbuk Province. The excysted metacercariae looked yellowish white and were $4.02{\pm}1.80$ mm (1.66-5.81) in length and $1.27{\pm}0.27$ mm (1.01-1.64) in width for acetocarmin-stained specimens. The oral sucker measuring $0.18{\pm}0.05$ mm (0.11-0.24) in length and $0.23{\pm}0.07$ mm (0.15-0.33) in width, was located at the anterior end of the body. The ventral sucker situated at the fourth third of the body were measured $0.47{\pm}0.08$ mm (0.35-0.53) in length and $0.55{\pm}0.09$ mm (0.43-0.63) in width. The tongue-shaped metacercariae were identified as Clinostomum complanatum on the basis of morphological characteristics. The metacercaria was also confirmed to be C. complanatum using PCR reaction with C. complanatum-specific primer sets, Trem 18S F and Ccom670 R. No histopathological changes in the infected fish were observed except necrosis and exfoliation of epidermis in the skin and atrophy of muscle fibers.

Identification of Monoecious and Dioecious Plants of. Schisandra nigra Using the RAPD Markers (RAPD 표지인자를 이용한 흑오미자의 자웅동주 및 자웅이주 식물의 동정)

  • 이효연;한효심;이갑연;한상섭;정재성
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.5
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    • pp.309-313
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    • 1998
  • RAPD (Random Amplified Polymorphic DNA) analysis was conducted to Schisandra nigra plants in order to select the specific markers for monoecious and dioecious individuals. RAPD results using eighty random 10-mer primers revealed that S. nigra had a different banding pattern from S. chinensis and Kadsura japonica. When DNA isolated from leaves of monoecious and dioecious plants were used as PCR template, only five primers, OPA-17, OPA-19, OPB-03, OPB-09 and OFB-16, showed polymorphic band patterns. No variation in banding profiles within male or female individuals was observed when these five primers were used whereas three monoecious plants (No 1, No 2 and No 3) showed different banding patterns one another, A 750 bp segment was amplified by primer OPB-3 from male individuals. On the other hand, two segments, 950 bp and 1690 bp, with OPA-19 and 700 bp of segment with OPB-3 were amplified in female individuals. These result indicate that the specific buds of male and female S. nigra could be used as genetic markers for the early discrimination of male and female individuals.

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Molecular Genetic Characterization and Analysis of Glucocorticoid Receptor Expression in the Big-belly Seahorse Hippocampus abdominalis (빅벨리해마(Hippocampus abdominalis) 글루코코르티코이드 수용체의 분자 유전학적 동정과 발현 분석)

  • Jo, Eunyoung;Oh, Minyoung;Lee, Sukkung;Qiang, Wan;Lee, Jehee
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.3
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    • pp.346-353
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    • 2015
  • Glucocorticoids (GCs) are steroid hormones regulated through responses to stress to maintain diverse metabolic and homeostatic functions. GCs act on the glucocorticoid receptor (GR), a member of the nuclear receptor family. This study identified and characterized the GR gene from the big-belly seahorse Hippocampus abdominalis designating it HaGR. The open reading frame of the HaGR cDNA was 2,346 bp in length, encoding a 782-amino-acid polypeptide with a theoretical isoelectric point of 6.26 and predicted molecular mass of 86.8 kDa. Nuclear receptors share a common structural organization, comprising an N-terminal transactivation domain, DNA-binding domain, and C-terminal ligand-binding domain. The tissue-specific mRNA expression profile of HaGR was analyzed in healthy seahorses using a qPCR technique. HaGR mRNA was expressed ubiquitously in all of the tissues examined, with the highest expression levels in kidney, intestine, stomach, and gill tissues. The mRNA expression in response to immune challenge with lipopolysaccharide (LPS), polyinosinic:polycytidylic acid (poly I:C), Edwardsiella tarda, and Streptococcus iniae revealed that it is inducible in response to pathogen infection. These results suggest that HaGR is involved in the immune response of the big-belly seahorse.

Yeasts Associated with Fruits and Blossoms Collected from Hanbat Arboretum, Daejeon, Korea (한밭 수목원의 과일과 꽃으로부터 효모의 분리)

  • Hyun, Se-Hee;Min, Jin-Hong;Kim, Seon-A;Lee, Jong-Soo;Kim, Ha-Kun
    • The Korean Journal of Mycology
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    • v.42 no.2
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    • pp.178-182
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    • 2014
  • Yeasts are common inhabitants of the phyllosphere, but our knowledge of their diversity in various fruits and blossoms are limited. We collected different kinds of blossoms and unripened fruits from Hanbat arboretum, Daejeon, Korea at the year of 2013. Yeasts were isolated by plating of suspensions prepared for collected samples onto YPD medium containing antibiotics. BLAST searches were subsequently performed for the comparison of the partially determined sequences of D1/D2 domain of 26S rDNA. As a result, we isolated 57 yeast strains of 31 species from 29 different kinds of blossoms and 6 kinds of fruits samples. We found huge differences in yeast flora depending on the sample collection season.

Isolation and Physiological Functionality of Yeasts from Wild Flowers in Seonyudo of Gogunsanyeoldo, Jeollabuk-do, Korea (전라북도 고군산열도의 선유도 일대 야생화들로부터 효모의 분리 및 생리기능성)

  • Hyun, Se-Hee;Han, Sang-Min;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.42 no.3
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    • pp.201-206
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    • 2014
  • Sixty one yeast strains of 21 species were isolated from wild flowers in Gogunsanyeoldo, including Seonyudo of Jeollabuk-do, Korea, and identified by comparison of nucleotide sequences for PCR-amplified D1/D2 region of 26S rDNA using BLAST. Among them, Cryptococcus sp. including C. aureus SY1-4 were found to be dominant, and Metschnikowia sp. including M. reukaufii SY20-1 and Rhodotnula sp. such as R. ingeniosa SY1-1 were also abundantly isolated. Some physiological functionalities of the culture broth and cell-free extracts from 61 yeast strains were determined. Supernatant from Metschnilowia reukaufii SY44-6 showed anti-gout xanthine oxidase inhibitory activity of 49.6% and whitening tyrosinase inhibitory activity of 38.4%, respectively.

Mitochondrial DNA Analysis of Human Skeletal Remains Excavated from Myungam-ri site in Asan, Korea (아산시 명암리 출토 인골의 미토콘드리아 DNA 분석)

  • Kim, Yun-Ji;Kim, Sue-Hoon;Cho, Eun-Min;Lee, Jeong-won
    • 보존과학연구
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    • s.36
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    • pp.33-48
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    • 2015
  • In this study, ancient DNA analyses were carried out on the human skeletal remains from a historical cemetery site in Myeongam-ri, Asan, Korea. Human remains of 27 individuals out of tombs from the Goryeo to Joseon Dynasty were selected for the analysis of this study. In order to identify the genealogy of the population and traditional burial pattern of the cemetery, we conducted comparative analyses of the hyper variable regions (HVRs) in mitochondrial DNA (mtDNA) of each sample. We sequenced 9 segmental amplicons of HVRs and assigned relevant haplogroups according to the sequence polymorphism on the basis of the known mtDNA database. As a result, we were analyses 18 human remains of 27 individuals and result of amelogenin analysis were only 4 samples.

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