• 제목/요약/키워드: rDNA sequence

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한국 토양으로부터 전분가수분해효소를 생산하는 고온성 균주의 선별과 동정 (Isolation and Characterization of Thermophilic Microorganism Producing Starch-hydrolyze Enzyme)

  • 최원석;배동훈
    • 산업식품공학
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    • 제14권1호
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    • pp.7-13
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    • 2010
  • 호열성 미생물을 검토하기 위하여 전국 각지로부터 토양과 두엄을 채취하여 그로부터 호열성 미생물을 분리하였다. 토양과 두엄으로부터 분리된 호열성 미생물 1250여 균주를 선별하였고, 이들을 대상으로 미생물이 생산하는 효소 활성을 검토하여 호열성 전분 분해 효소를 생산하는 1주의 미생물을 확인하였다. 확인된 1주의 미생물을 strain 2719라 명명하였다. Strain 2719 균주는 형태학적으로 gram 양성 간균의 특징을 나타냈고, 균주의 표면은 매끄럽지 않았으며, 비교적 다양한 길이를 가지고 있었다. 또한 다른 gram 양성 간균들에 비해서 많은 수의 균사들이 각 균주들 사이에 복잡하게 얽혀있었다. 생화학적 특성을 확인한 결과 catalase 양성, glucose 발효, arabinose 발효, mannitol 발효, casein gelatin starch 가수분해의 특징을 가지고 있었으며, 이는 Bacillus sp.로 추정되었다. 생육 pH의 범위는 pH 6-pH 8범위에서 생육이 가능했으며, 생육 온도의 범위는 50-70${^{\circ}C}$였다. 16S rDNA sequence 분석결과 Bacillus thermoglucosidasius의 16S rDNA와 99.52%가 일치하였으나, sequence의 일부분이 다른 부분이 있고, 생육 특성에서 약간의 차이를 보였다. 또한 gene bank에 등록되어 있는 균주들의 16S rDNA sequence들과 비교하여도 일치하는 균주는 확인되지 않았다. 이와 같은 실험결과에 따라 2719 균주는 기존에 발표되지는 않았으나, Bacillus thermoglucosidasius와 매우 유사한 균주로 판단되어 Bacillus thermoglucosidasius 2719로 명명하였다.

동해 연안에 서식하는 성게의 형태변이와 미토콘드리아 유전자 분석 (Morphological Variation and Partial Mitochondrial Sequence Analysis of Echinoid Species from the Coasts of the East Sea)

  • 신지혜;김성규;김영대;손영창
    • 한국양식학회지
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    • 제21권3호
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    • pp.139-145
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    • 2008
  • 성게의 형태학적 분류는 그것의 형질적 변이에 의하여 어려움이 많다. 본 연구에서는 말똥성게, 둥근성게, 보라성게, 분홍성게와 동해안에서 포획된 미확인 성게 4종의 형태형질 비교와 계통유연관계를 조사하였다. 성게의 생식소로부터 genomic DNA를 분리한 후, PCR 방법을 통하여 mitochondrial 12S rDNA 유전자의 염기서열을 분석하였다. 둥근성게과의 말똥성게, 둥근성게, 만두성게과의 보라성게, 주발성게과의 분홍성게의 mitochondrial 12S rDNA의 염기서열은 미확인 성게종들의 그것과 85.9-93.9%의 상동성을 나타내었다. 한편, 미확인 성게종들은 새치성게의 mitochondrial 12S rDNA의 일부 염기서열과 99.8%의 높은 상동성을 보였으며, 각 개체의 mitochondrial 12S rDNA를 통한 분자계통수 분석에 의해서 미확인 성게들은 새치성게의 형태적 변이로 판단된다.

Sequence comparisons of 28S ribosomal DNA and mitochondrial cytochrome c oxidase subunit I of Metagonimus yokogawai, M. takahashii and M. miyatai

  • Lee, Soo-Ung;Huh, Sun;Sohn, Woon-Mok;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • 제42권3호
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    • pp.129-135
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    • 2004
  • We compared the DNA sequences of the genus Metagonimus: M. yokogawai, M. takahashii, and M. miyatai. We obtained 288 D1 ribosomal DNA (rDNA) and mitochondrial cytochrome c oxidase subunit I (mtCOI) fragments from the adult worms by PCR, that were cloned and sequenced. Phylogenetic relationships inferred from the nucleotide sequences of the 28S D1 rDNA and mtCOI gene. M. takahashii and M. yokogawai are placed in the same clade supported by DNA sequence and phylogenie tree analysis in 28S D1 rDNA and mtCOI gene region. The above findings tell us that M. takahashii is closer to M. yokogawai than to M. miyatai genetically. This phylogenetic data also support the nomination of M. miyatai as a separate species.

Characterization of Trichoderma spp. Associated with Green Mold of Oyster Mushroom by PCR-RFLP and Sequence Analysis of ITS Regions of rDNA

  • Park, Myung-Soo;Seo, Geon-Sik;Bae, Kyung-Sook;Yu, Seung-Hun
    • The Plant Pathology Journal
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    • 제21권3호
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    • pp.229-236
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    • 2005
  • Molecular profIles of PCR-RFLP and sequence analysis of internal transcribed spacer (ITS) regions of rDNA were compared between morphologically distinguishable species of Trichoderma isolated from substrates of oyster mushroom in Korea, T. atroviride, T. citrinoviride, T. harzianum, T. longibrachiatum, T. virens, and two unidentified species, Trichoderma sp. 1 and 2. PCR­RFLP analysis divided the Trichoderma spp. into six RFLP groups, A, B, C, D, E, and F. The RFLP groups were generally agreed with described morphological species, except that the RFLP group A containing the two unidentified species. A neighbor-joining tree based on ITS sequences well supported RFLP groups observed by RFLP analysis of ITS regions of rDNA. Additionally, the two unidentified species, Trichoderma sp. 1 and 2, which could not be distinguished by PCR­RFLP analysis, were separated in sequence analysis of ITS regions of rDNA.

Sequence Comparison of Mitochondrial Small subunit Ribosomal DNA in Penicillium

  • Bae, Kyung-Sook;Hong, Soon-Gyu;Park, Yoon-Dong;Wonjin Jeong
    • Journal of Microbiology
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    • 제38권2호
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    • pp.62-65
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    • 2000
  • Partial sequence comparisons of mitochondrial small subunit rDNA (mt SSU rDNA) were used to examine taxonomic and evolutionary relationships among seven Penicillium species : two monoverticillate species, two biverticillate species, and three terverticillate species. Amplified fragments of mt SSU rDNA highly varied among seven species in size, suggesting the existence of multiple insertions or deletions in the region. A phylogengtic tree was constructed by exhaustive search of parsimony analysis. The phylogenetic tree distinguished two statistically supported monophyletic groups, one for two monoverticillate species and the other for three terverticillate species and ont biverticillate species, P. vulpinum. The phylogenetic relationship of P. waksmanii, the biverticillate species, was not clear.

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Physiological and Phylogenetic Analysis of Burkholderia sp. HY1 Capable of Aniline Degradation

  • Kahng, Hyung-Yeel;Jerome J. Kukor;Oh, Kye-Heon
    • Journal of Microbiology and Biotechnology
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    • 제10권5호
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    • pp.643-650
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    • 2000
  • A new aniline-utilizing microorganism, strain HY1 obtained from an orchard soil, was characterized by using the BIOLOG system, an analysis of the total cellular fatty acids, and a 16S rDNA sequence. Strain HY1 was identified as a Burkholderia species, and was designated Burkholderia sp. HY1. GC and HPLC analyses revealed that Burkholderia sp. HY1 was able to degrade aniline to produce catechol, which was subsequently converted to cis,cis-muconic acid through an ortho-ring fission pathway under aerobic conditions. Strain HY1 exhibited a drastic reduction in the rate of aniline degradation when glucose was added to the aniline media. However, the addition of peptone or nitrate to the aniline media dramatically accelerated the rate of aniline degradation. A fatty acid analysis showed that strain HY1 was able to produce lipids 16:0 2OH, and 11 methyl 18:1 ${\omega}7c$ approximately 3.7-, 2.2-, and 6-fold more, respectively, when grown on aniline media than when grown on TSA. An analysison the alignment of a 1,435 bp fragment. A phylogenetic analysis of the 16S rDNA sequence based on a 1,420 bp multi-alignment sowed of the 16s rDNA sequence revealed that strain HY1 was very closely related to Burkholderia graminis with 95% similarity based that strain HY1 was placed among three major clonal types of $\beta$-Proteobacteria, including Burkholderia graminis, Burkholderia phenazinium, and Burkholderia glathei. The sequence GAT(C or G)${\b{G}}$, which is highly conserved in several locations in the 16S rDNA gene among the major clonal type strains of $\beta$-Proteobacteria, was frequently replaced with GAT(C or G)${\b{A}}$ in the 16S rDNA sequence from strain HY1.

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Nuclear DNA Quantification of Some Ceramialean Algal Spermatia by Fluorescence Microscopic Image Processing and their Nuclear SSU rDNA Sequences

  • Choi, Han-Gu;Lee, Eun-Young;Oh, Yoon-Sik;Kim, Hyung-Seop;Lee, In-Kyu
    • ALGAE
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    • 제19권2호
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    • pp.79-90
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    • 2004
  • Nuclear DNA contents of spermatia from eight ceramiacean and four dasyacean algae (Ceramiales, Rhodophyta) and microspores from two land plants were estimated by fluorescence microscopic image processing and their nuclear SSU rDNA sequence data were analyzed. In frequency distribution patterns, the DAPI-stained nuclear volume (NV) of spermatia showed two peaks corresponding to 1C and 2C. Nuclear 2C DNA contents estimated from NV were 0.45-2.31 pg in ceramiacean and 0.40-0.57 pg in dasyacean algae and 8.42-9.51 pg in two land plants, Capsicum annuum and Nicotiana tabacum. By nuclear patterning of vegetative cells derived from an apical cell, 2C DNA contents of spermatia were 2.31 pg in an alga having uninucleate and non-polyploid nucleus (Aglaothamnion callophyllidicola), 0.45-1.94 pg in algae having uninucleate and polyploid nucleus (Antithamnion spp. and Pterothamnion yezoense), and 0.40-0.62 pg in algae having multinucleate and non-polyploid nuclei (Griffithsia japonica and dasyacean algae). Each mature spermatium and microspore (pollen grain) seemed to have a 2C nucleus, which may provide a genetic buffering system to protect the genetic content of a spermatium and microspore from potentially lethal mutations. Nuclear DNA content and SSU rDNA sequence of Antithamnion sparsum from Korea were reasonably different from those of Antithamnion densum from France. The data did not support the previous taxonomic studies that these two taxa could be conspecific.

Characterization of Sclerotinia sclerotiorum Isolated from Paprika

  • Jeon, Young-Jae;Kwon, Hyuk-Woo;Nam, Ji-Sun;Kim, Seong-Hwan
    • Mycobiology
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    • 제34권3호
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    • pp.154-157
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    • 2006
  • A fungal isolate collected from infected paprika (Capsicum annuum var. grossum) was characterized as Sclerotinia sclerotiorum based on its ability of sclerotium formation, physiological and molecular properties. When the isolate was grown on potato dextrose agar, oatmeal agar, and malt extract agar, it grew most well on PDA. Optimal temperature and pH for its growth were $25^{\circ}C$ and pH 7, respectively. The fungal isolate produced sclerotia on PDA within 10 days, and the color and shape of the sclerotia were similar to those of S. sclerotiorum. The ITS rDNA regions including ITS1 and ITS2 and 5.8S sequences were amplified using ITS1F and ITS4 primers from the genomic DNAs of the paprika isolate and other known pathogenic S. sclerotiorum isolated from different crops in Korea, and their nucleotide sequences were determined. Sequence comparison analysis showed the ITS rDNA of the paprika isolate shares 100% sequence identity with those of S. sclerotiorum isolated from red pepper, lettuce and a S. sclerotiorum isolate registered in GenBank DNA database. Neighbor joining analysis based on the ITS rDNA sequence revealed the paprika isolate has very close phylogenetic relationships with known Sclerotinia sclerotiorum isolates. This is the first report that S. sclerotiorum has been found associated with paprika rot in paprika growing countries.

Cloning and Characterization of the Putative Transferrin Receptor cDNA from the Olive Flounder (Paralichthys olivaceus)

  • Won Kyoung-Mi;Park Soo-Il
    • Fisheries and Aquatic Sciences
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    • 제6권2호
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    • pp.101-104
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    • 2003
  • A cDNA clone for the olive flounder (Paralichthys olivaceus) transferrin receptor (fTfR) was isolated from a leukocytes cDNA library. The fTfR gene consisted of 2,319 bp encoding 773 amino acid residues. The amino acid sequence alignment of the fTfR showed that their size and hydrophobic profile are similar. In addition, the Tyr-Thr-Arg-Phe (YTRF) motif that is the recognition signal for high-efficiency endocytosis, is conserved very well. This motif is important for functional properties of TfR. The deduced amino acid sequence had $42.4-42.9\%$ identities with the previously reported TfRs of vertebrates. The fTfR was expressed in the blood, kidney, spleen, and liver of healthy olive flounder by the Northern blot hybridization.

Phylogenetic Relationships of the Aphyllophorales Inferred from Sequence analysis of Nuclear Small Subunit Ribosomal DNA

  • Kim, Seon-Young;Jung, Hack-Sung
    • Journal of Microbiology
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    • 제38권3호
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    • pp.122-131
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    • 2000
  • Phylogenetic classification of the Aphyllophorales was conducted based on the analysis of nuclear small subunit ribosomal RNA (nuc SSU rDNA) sequence. Based on phylogenetic groupings and taxonomic characters, 16 families were recognized and discussed. Although many of the characters had more or less homoplasies, miroscopic characters such ad the mitic system and clamp, spore amyloidity and rot type appeared to be important in the classification of the Aphyllophorales. Phylogenetically significant families were newly defined to improve the classification of the order Aphyllophorales.

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