• Title/Summary/Keyword: ITS rDNA

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Detection of DNA Sequence Polymorphism by Polymerase Chain Reaction in Fraxinus mandshurica Rupr Growing in Korea (P.C.R 기법(技法)을 이용(利用)한 들메나무 DNA sequence의 변이조사(變異調査))

  • Na, Chun Soo;Noh, Eun Woon;Kim, Young Joong;Shin, Chang Ho;Song, Won Sup;Kim, Sea Hyun
    • Journal of Korean Society of Forest Science
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    • v.81 no.4
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    • pp.320-324
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    • 1992
  • It has been reported that there are two distinct phenotypes in Fraxinus mandshurica Rupr. growing in Korea. Recently developed polymerase chain reaction(PCR) was used to detect DNA sequence polymorphism in the species. Using a thermostable DNA polymerase and synthetic DNA primers, unknown DNA sequences from the species were randomly amplified. The two types of the species produced different DNA amplification pattern with three different primers tested. Although DNA polymorphism was detected among individuals within types, each type has its own distinct pattern. The two types could be easily differentiated by trier characteristic predominant bands.

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Quantitative Detection of Residual E. coli Host Cell DNA by Real-Time PCR

  • Lee, Dong-Hyuck;Bae, Jung-Eun;Lee, Jung-Hee;Shin, Jeong-Sup;Kim, In-Seop
    • Journal of Microbiology and Biotechnology
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    • v.20 no.10
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    • pp.1463-1470
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    • 2010
  • E. coli has long been widely used as a host system for the manufacture of recombinant proteins intended for human therapeutic use. When considering the impurities to be eliminated during the downstream process, residual host cell DNA is a major safety concern. The presence of residual E. coli host cell DNA in the final products is typically determined using a conventional slot blot hybridization assay or total DNA Threshold assay. However, both the former and latter methods are time consuming, expensive, and relatively insensitive. This study thus attempted to develop a more sensitive real-time PCR assay for the specific detection of residual E. coli DNA. This novel method was then compared with the slot blot hybridization assay and total DNA Threshold assay in order to determine its effectiveness and overall capabilities. The novel approach involved the selection of a specific primer pair for amplification of the E. coli 16S rRNA gene in an effort to improve sensitivity, whereas the E. coli host cell DNA quantification took place through the use of SYBR Green I. The detection limit of the real-time PCR assay, under these optimized conditions, was calculated to be 0.042 pg genomic DNA, which was much higher than those of both the slot blot hybridization assay and total DNA Threshold assay, where the detection limits were 2.42 and 3.73 pg genomic DNA, respectively. Hence, the real-time PCR assay can be said to be more reproducible, more accurate, and more precise than either the slot blot hybridization assay or total DNA Threshold assay. The real-time PCR assay may thus be a promising new tool for the quantitative detection and clearance validation of residual E. coli host cell DNA during the manufacturingprocess for recombinant therapeutics.

Cloning of Steroid $\Delta^1$-dehydrogenase Gene of Arthrobacter simplex IAM 1660

  • Bae, Moo;Bae, Song-Mee;Lee, Mi-Kyung;Lee, Jeong-Kug
    • Journal of Microbiology and Biotechnology
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    • v.6 no.2
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    • pp.142-144
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    • 1996
  • To clone the gene coding for steroid $\Delta^1$-dehydrogenase of Arthrobacter simplex, its genomic library was constructed with a , $\lambda$gt11 expression vector and immunoscreened with antiserum against the enzyme. One positive clone was found to carry a 1.6-kb EcoR I restriction endonuclease fragment of A. simplex DNA. The restriction map of the 1.6-kb EcoR I fragment was determined after cloning of the DNA into pBS vector.

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Axenic Culture of Gyrodinium impudicum Strain KG03, a Marine Red-tide Microalga that Produces Exopolysaccharide

  • Yim Joung Han;Lee Hong Kum
    • Journal of Microbiology
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    • v.42 no.4
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    • pp.305-314
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    • 2004
  • An exopolysaccharide-producing microalgal dinoflagellate was isolated from a red-tide bloom and des­ignated strain KG03. A bacteria-free culture of strain KG03 was achieved using a modified wash with phototaxis and antibiotic treatment. Combined treatment with neomycin and cephalosporin was the most effective for eliminating the bacteria associated with the microalgae. Strain KG03 was identified as Gyrodinium impudicum by analyzing the ITS regions of the 5.8S rDNA, 18S rDNA, morphological phenotype and fatty acid composition. The exopolysaccharide production and cell growth in a 300-ml photobioreactor were increased 2.7- and 2.4-fold, respectively, compared with that in a flask culture at the first isolation step.

Production of Bacterial Cellulose by Gluconacetobacter hansenii PJK Isolated from Rotten Apple

  • Park, Joong-Kon;Park, Youn-Hee;Jung, Jae-Yong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.2
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    • pp.83-88
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    • 2003
  • A cellulose-producing strain isolated from rotten apples was identified as Gluconacetobacter hansenii based on its physiological properties and the 16S rDNA complete sequencing method, and specifically named Gluconacetobacter hansenii PJK. The amount of bacterial cellulose (BC) produced by G. hansenii PJK in a shaking incubator was 1.5 times higher than that produced in a static culture. The addition of ethanol to the medium during cultivation enhanced the productivity of bacterial cellulose, plus the supplementation of 1% ethanol into the culture medium made the produced BC aggregate into a big lump and thus protected the bacterial-cellulose-producing G. hansenii PJK cells in the shear stress field from being converted into non-cellulose-producing (Cel) mutants. Cells subcultured three times in a medium containing ethanol retained their ability to produce BC without any loss in the production yield.

Two New Species of Cryptococcus sp. and Candida sp. from Wild Flowers in Korea

  • Min, Jin-Hong;Kang, Min-Gu;Ryu, Jin-Ju;Lee, Hyang-Burm;Kim, Chang-Mu;Kim, Ha-Kun;Lee, Jong-Soo
    • Mycobiology
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    • v.40 no.4
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    • pp.255-257
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    • 2012
  • Among 80 types of yeast isolated from wild flowers in Daejeon, Korea, two species that have not yet been identified by phylogenetic analysis of the internal transcribed spacer-2 (ITS2) genes and 26S rDNA sequences were identified as Candida sp. 44-C-1 and Cryptococcus sp. 9-D-1. Neither of the newly identified species formed ascospores, while Candida sp. 44-C-1 formed pseudomycelium and Cryptococcus sp. 9-D-1 did not.

The First Report of Antrodia sitchensis (Polyporaceae, Basidiomycota) in Korea

  • Jang, Yeong-Seon;Choi, Ha-Eun;Lim, Young-Woon;Lee, Jin-Sung;Kim, Jae-Jin
    • Mycobiology
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    • v.39 no.3
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    • pp.226-229
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    • 2011
  • An unrecorded Antrodia species was collected in South Korea and based on morphological characteristics, the species was identified as Antrodia sitchensis. To confirm its affinity within the polypores, the phylogenetic relationships of A. sitchensis and allied species were established using large subunit rDNA sequences.

New Records of Three Tintinnopsis Species and Redescription of One Antetintinnopsis Species (Protozoa: Ciliophora) from Coastal Waters of Korea

  • Ji Hye Moon;Jae-Ho Jung
    • Animal Systematics, Evolution and Diversity
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    • v.40 no.1
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    • pp.1-15
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    • 2024
  • Our study aimed to investigate the diversity of tintinnid species in Korea by collecting samples from coastal waters. As a result, we identified and redescribed three newly recorded species of the genus Tintinnopsis Stein, 1867 and one previously recorded species of the genus Antetintinnopsis Wang et al., 2021 in Korea. The loricae morphology and molecular phylogeny based on the 18S rDNA sequences of these four were analyzed. Tintinnopsis kiaochowensis Yin, 1956 is characterized by having an irregular collar with spiral turns and an obconical-shaped bowl. Tintinnopsis orientalis Kofoid and Campbell, 1929 is characterized by the inverted bell-shaped lorica with size of 121-140×86-94 ㎛. Tintinnopsis parvula Jörgensen, 1912 is characterized by its narrower collar than bowl and acute angle of the bowl (39-75°). The recorded species, Antetintinnopsis gracilis (Kofoid and Campbell, 1929) Wang et al., 2021 is characterized by a cylindrical collar narrower than the bowl width and with a size of 98-131×37-46 ㎛.

Cohesion Establishment Factors Stimulate Endonuclease Activity of hFen1 Independently and Cooperatively

  • Kim, Do-Hyung;Kim, Jeong-Hoon;Park, Byoung Chul;Cho, Sayeon;Park, Sung Goo
    • Journal of Microbiology and Biotechnology
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    • v.25 no.10
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    • pp.1768-1771
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    • 2015
  • Human Fen1 protein (hFen1) plays an important role in Okazaki fragment processing by cleaving the flap structure at the junction between single-stranded (ss) DNA and doublestranded (ds) DNA, an intermediate formed during Okazaki fragment processing, resulting in ligatable nicked dsDNA. It was reported that hChlR1, a member of the cohesion establishment factor family, stimulates hFen1 nuclease activity regardless of its ATPase activity. In this study, we found that cohesion establishment factors cooperatively stimulate endonuclease activity of hFen1 in in vivo mimic condition, including replication protein-A-coated DNA and high salt. Our findings are helpful to explain how a DNA replication machinery larger than the cohesion complex goes through the cohesin ring structure on DNA during S phase in the cell cycle.

Analysis of the Role of RGG box of human hnRNP A1 protein (인간 hnRNP A1 단백질에 포함된 RGG 상자의 기능 분석)

  • Choi, Mieyoung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.12
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    • pp.575-580
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    • 2017
  • This study analyzed the effects of RGG box of hnRNP A1 on its subcellular localization and stabilization of hnRNP A1 over a three year period from October 2014. First, a 6R/K mutation in RGG box was generated, and pcDNA1-HA-hnRNP A1(6R/K) was constructed. The subcellular localization of hnRNP A1(6R/K) from the HeLa cells transfected with this plasmid DNA was analyzed by immunofluorescence microscopy. HA-hnRNP A1(6R/K) was found to exhibit nuclear and cytoplasmic fluorescence. The stability of hnRNP A1(6R/K) was checked by Western blot analysis using the expressed protein from the HeLa cells transfected with the pcDNA1-HA-hnRNP A1(6R/K). The results show that HA-hnRNP A1(6R/K) has a smaller size. These confirm that HA-hnRNP A1(6R/K) is localized both in the nuclear and cytoplasm, not because 6R/K mutation affects the nuclear localization of hnRNP A1, but because 6R/K mutation causes hnRNP A1(6R/K) to cleave at the mutation or near the mutation site. The cleaved protein fragment, which lacks the M9 domain (i.e. nuclear localization signal of hnRNP A1), did not exhibit nuclear fluorescence. This suggests that the arginines of RGG box in hnRNP A1 play an important role in stabilizing hnRNP A1. An analysis of the RNA-binding ability of hnRNP A1(6R/K) expressed and purified from bacteria will be a subsequent research project.