• Title/Summary/Keyword: 45S and 5S rDNA

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Antibacterial Effect of Bacteria Isolated from the Korean Traditional Foods against Pathogenic Bacteria (한국전통식품으로부터 분리 된 세균의 항균활성 효과)

  • Moon, Kyung-Mi;Heo, Moon-Soo
    • Journal of Life Science
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    • v.25 no.11
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    • pp.1319-1323
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    • 2015
  • Aquaculture continues to be an ever-growing sector. However, high-density farming increases disease outbreaks due to deteriorating water quality and internal stress. To prevent disease, the most common method chemotherapy is using antibiotic administration. In this study, probiotic bacteria were isolated from Korean traditional foods, such a Gochu pickle and cutlassfish salted seafood. Various bacteria were isolated, and their 16S rDNA sequences were analyzed. The antimicrobial activities of four isolates from Gochu pickle and seven isolates from cutlassfish salted seafood were assayed, in addition to the antibacterial activity of culture pellet and supernatant. The antibacterial activity of the pellet was higher than that of the supernatant. Isolate JKM-2 showed the highest antibacterial activity against Streptococcus iniae (43 mm), S. parauberis (40 mm), S. mutans (35 mm), and Vibrio vuinificus (26.5 mm). The sequences of the isolated strains were compared with those of Bacillus subtilis (97.71%), B. tequilensis (97.71%), Brevibacterium halotolerans (97.71%), B. subtilis (97.63%), B. subtilis (97.63%), B. mojavensis (97.54%), B. vallismortis (97.46%), B. nanillea (97.45%), B. methylotrophicus (97.37%), and B. ssiamensis (97.37%). Future through analysis and new strains confirmed the bacterial cell material investigation of JKM-3, and to ensure sufficient stability, it is desired to verify the utility value as a substitute material for antibiotics by application to the form of the industry.

Occurrence of Plant Parasitic Nematodes in Chrysanthemum and ITS and D3-28S rDNA Characterization of Pratylenchus spp. (국화재배지의 식물기생선충 분포조사 및 뿌리썩이선층의 ITS와 D3-28S rDNA 특성조사)

  • Han, Hye-Rim;Lee, Jae-Kook;Choi, Dong-Ro;Han, Man-Jong;Park, Byeong-Yong
    • Korean journal of applied entomology
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    • v.45 no.3 s.144
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    • pp.293-299
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    • 2006
  • A survey was conducted to find out the major plant parasitic nematode in Chrysanthemum morifolium fields in Korea from May to June in 2005. A genus of Pratylenchus was determined as the most important plant parasitic nematode based on analysis of total 50 samples from 8 cities of chrysanthemum field. Pratylenchus showed 86% occurrence rate and average numbered 1,095 per 200cc soils and 1g root. Five Pratylenchus isolates, 'Muan', 'Masan', 'Tean', 'Gumi', 'Jeongup', were selected for the molecular identification of the species of Pratylenchus, and ITS and D3-28S ribosomal DNA were amplified by PCR. For the ITS, only 'Muan' isolate was differentiated by total 1 kb PCR amplification, which was 200 bp larger than all the other isolates. There was no size variation in amplified D3-28S rDNA and all isolate represented approximately 320 bp of PCR product. Sequence data of D3-28S rDNA were analysed by MegAlign program in DNASTAR software and phylogenetic tree was constructed. Sequence homology was 100% between 'Gumi' isolate and 'Tean' isolate and 'Jeongup' isolate was also close to these isolates by 99.7% sequence homology. 'Gumi', 'Tean' group and 'Jeongup' isolate were determined to be closely related to Pratylenchus vulnus by 96.7% and 96.3% similarity in respectively. D3 sequence of 'Masan' isolate was 100% identical to P. penetrans, and 'Muan' isolate showed 99.7% similarity to P. brachyurus. This result was congruent with the branch divergence pattern shown in phylogenetic tree.

Comparison of Bacterial Composition between Human Saliva and Dental Unit Water System

  • Jeon, Eun-Hyoung;Han, Ji-Hye;Ahn, Tae-Young
    • Journal of Microbiology
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    • v.45 no.1
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    • pp.1-5
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    • 2007
  • The bacterial compositions between the dental unit water system and human saliva were characterized and compared by direct sequence analysis of 16S rDNA clone libraries. Based on the species richness estimation, bacterial diversity in the dental unit water system (DUW) was more diverse than that of the human saliva (HS). The Chaol estimates of species richness in HS and DUW samples were 12.0 and 72.4, respectively. The total numbers of OTUs observed in the combined libraries accounted for 83% (HS) and 59% (DUW) of the Chaol diversity estimate as defined at the 80% similarity threshold. Based on the sequence analysis, the phylum Proteobacteria was the major group in both clone libraries at phylum level. DUW clone library contained 80.0% Proteobacteria, 8.0% Bacteroides, 4.0% Nitrospira, 4.0% Firmicutes, 2.0% Planctomycetes and 2.0% Acidobacteria. On the other hand, human saliva (HS) clone library contained 55.5% Proteobacteria, 36.1% Firmicutes and 8.4% Bacteroides. The majority of bacteria identified belonged to phylum Proteobacteria in both samples. In dental unit water system (DUW), Alphaproteobacteria was detected as the major group. There was no evidence of the bacterial contamination due to a dental treatment. Most sequences were related to microorganisms derived from biofilm in oligotrophic environments.

Molecular Cloning and Characterization of a Flavanone-3-hydroxylase Gene from Rubus occidentalis L.

  • Lee, Seung Sik;Lee, Eun Mi;An, Byung Chull;Barampuram, Shyamkumar;Kim, Jae-Sung;Cho, Jae-Young;Lee, In-Chul;Chung, Byung Yeoup
    • Journal of Radiation Industry
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    • v.2 no.3
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    • pp.121-128
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    • 2008
  • Flavanone-3-hydroxylase (F3H) is one of the key enzymes for the biosynthesis of flavonals, anthocyanins, catechins and proanthocyanins. F3H catalyzes the $3{\beta}$-hydroxylation of (2S)-flavonones to form (2R, 3R)-dihydroflavonols. In this report, we isolated a full-length cDNA of RocF3H from black raspberry (Rubus occidentalis L.) using a reverse transcriptase-PCR and rapid amplification of the cDNA ends (RACE)-PCR. The full-length cDNA of RocF3H contains a 1,098 bp open reading frame (ORF) encoding a 365 amino acid protein with a calculated molecular weight of about 41.1 kDa and isoelectric point (pI) of 5.45. The genomic DNA analysis revealed that the RocF3H gene had three exons and two introns. Comparison of the deduced amino acid sequence of the RocF3H with other F3Hs revealed that the protein is highly homologous with various plant species. The conserved amino acids ligating the ferrous iron and the residues participating in the 2-oxoglutarate binding (R-X-S) were found in RocF3H at the similar positions to other F3Hs. Southern blot analysis indicated that RocF3H exist a multi-gene family. The isolation of RocF3H gene will be helpful to further study the role of F3H gene in the biosynthesis of flavonoids in R. occidnetalis.

Occurrence of Sporendocladia bactrospora on Quercus variabilis in Korea

  • Lee, Dong-Hyeon;Seo, Sang-Tae;Lee, Sang-Hyun;Lee, Seung Kyu;Lee, Sun Keun
    • The Korean Journal of Mycology
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    • v.45 no.4
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    • pp.394-398
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    • 2017
  • A survey to assess the diversity of wound-associated Ophiostomatales and Microascales, the so-called ophiostomatoid fungi, on Korean native trees, was undertaken in 2017. Wounds were artificially created, and a fungus resembling a species of Sporendocladia was consistently isolated from the exposed cambium and inner bark of artificially induced wounds on Quercus variabilis. Morphological examination and DNA sequence comparisons based on the internal transcribed spacer (ITS) and 5.8S regions of the rDNA confirmed the identity of the fungus as Sporendocladia bactrospora. This is the first report on S. bactrospora occurring on Q. variabilis in Korea.

Studies on Saprogenic Bacteria Isolated from Korean Red Ginseng Extrart Product (홍삼 추출액 파우치 제품으로부터 분리동정된 변패세균에 관한 연구)

  • Kim, Young-Man;Yoon, Choong-Eui;Kim, Kyoung-Hee;Lee, Eun-Woo
    • Journal of Life Science
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    • v.17 no.10
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    • pp.1394-1399
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    • 2007
  • Five kinds of saprogenic bacteria were isolated from the red ginseng extract product and identified as Bacillus subtilis, Bacillus sp., Paenibacillus sp., Micrococcus sp., and Pseudomonas sp. by 16S rDNA analysis. Some of the isolated strains were able to grow even at $45^{\circ}C$ which are presumed originated from the raw ingredient of red ginseng extract. All of the isolated strains did not show the hemolytic activity, the diarrhea-inducing activity, and the vascular permeability enhancing activity, indicating that these strains are not pathogenic.

Genetic Study of the Class Dinophyceae Including Red Tide Microalgae Based on a Partial Sequence of SSU Region : Molecular Position of Korean Isolates of Cochlodinium polykrikoides Margalef and Gyrodinium aureolum Hulburt (SSU 부위의 유전자 염기서열 분석에 의한 한국연안에서 분리한 Cochiodinium polykrikoides Margalef와 Gyrodinium aurelum Hulburt 적조생물의 분자생물학적 연구)

  • Cho, Eun-Seob
    • Journal of Life Science
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    • v.14 no.4
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    • pp.593-607
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    • 2004
  • The nucleotide sequence for a nuclear-encoded small subunit rDNA (SSU rDNA) was determined for 43 species of the class Dinophyceae, including harmful algae Cochlodinium polykrikoides and Gyrodinium aureolum. These sequences and data analyses were performed by parsimony, distances and maximum likelihood methods in PHYLIP (Phylogenetic Inference Package) version 3.573c. The species Noctiluca scintillans, Gonyaulax spinifern and Crypthecodinium cohnii occupied a basal position within the Dino- phyceae in our analyses. The genera Alexandrium and Symbiodinium were monophyletic (supported by a bootstrap value of >70%), whereas the genera Gymnedinium and Gyrodinium formed polyphyletic nodes, for which bootstrap support was strong (>70%) in the neighbor-joining and maximum likelihood methods except for the PHYLIP parsimony analysis (=59%). The sequence divergence between G. aureolum and G. dorsum/ G. galathenum was the largest at 7.4% (45 bp), whereas G. aureolum and G. mikimotoi showed an extremely low value of genetic divergence of 0.9% (5 bp). The genetic divergence between C. polykrikoides and G. aureolum was a low value of 5.2% (31 bp). In the phylogenetic analysis, the placement of G. aureolum and C. polykrikoides was closer to the genus Gymnodinium than to the genus Gyrodinium, which was supported by a moderate bootstrap value.

Characterization of Ovarian Cytochrome $P450_{C17}$ (17 ${\alpha}-hydroxylase$/17,20-lyase) in Rana dybowski (북방산 개구리 난소의 Cytochrome $P450_{C17}$ 유전자 특성)

  • Kang, Hae-Mook
    • Development and Reproduction
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    • v.10 no.2
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    • pp.127-133
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    • 2006
  • [ $17\;{\alpha}-hydroxylase/17,20-lyase(P450_{C17})$ ] is the key enzyme mediating the conversion of progesterone to $17\;{\alpha}-hydroxyprogesterone$, ultimately to androstenedione during steroidogenesis. R. dybowskii's ovarian $P450_{C17}$ cDNA was cloned to understand the regulatory mechanism of ovarian steroidogenic pathway at the molecular level in amphibian. A 2.5kb cDNA clone encoding a single open-reading frame with a 519 deduced amino acid was isolated with the screening of ovarian cDNA library. This sequence contained the three highly conserved domains as seen in $P450_{C17}$ of other species. The comparison of amino acid sequence of Rana $P450_{C17}$ with other animal's $P450_{C17}$ showed relatively high identity with 76% in Xenopus, 63% in chicken, 60% in rainbow trout, and 45% in human. Phylogenic analysis also indicated that Rana $P450_{C17}$ gene was evolutionary well conserved among vertebrate. Northern analysis indicated that the two different sizes of $P450_{C17}$ transcripts with approximately 2.5 and 3.6kb were detected in ovary tissue, but not in other tissues. The expression vector of Rana $P450_{C17}$ clearly showed the $17\;{\alpha}-hydroxylase$ activity converting the exogenous progesterone into $17\;{\alpha}-hydroxyprogesterone$ in the nonsteroidogenic COS-1 cells. Therefore, Rana $P450_{C17}$ cDNA is very useful to investigate the molecular mechanism of the ovarian steroidogenesis in amphibian.

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Isolation and Characterization of Mitochondrial DNA from Arehicityars and Metacitrus (Archicitrus와 Metacitrus로부터 Mitochondrial DNA의 분리 및 특성)

  • 이숙영;박민희
    • The Korean Journal of Food And Nutrition
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    • v.8 no.4
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    • pp.307-317
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    • 1995
  • The purity of mtDNAs isolated from Archicitrus and Metacitrus leaves was higher in percoll density gradient centrifugation than differential and sucrose density gradient centrifugation. The most clear mtDNAs were obtained from mitochondria included in the Interface band of between 21% and 45% under isomotic, low viscosity conditions in the three step discontinuous percoll density gradient centrifugation. DNase treatment to the crude mitochondrlal suspension still more increased purity of mtDNA by the effective removal of the nuclear and chloroplast DNA and mtDNAs were appeared as a single band at middle position of tube by EtBr /cscl density gradient centrifugation. Agarose gel electrophoresis of mtDNAs resolved a single, broad band containing high molecular weight DNAs In all preparation. Yield of mtDNAs was about 110 and 2 ug Per 2009 in mature and immature leaves respectively. The mtDNA fragment patterns showed by EcoR I treatment were indistinguishable with respect to nom bet and position of bands in Archicitrus and Metacitrus. In the pattern of Hind E restriction, the Metacitrus displayed the unique band between 5.0 and 4.0kb, in addition to four fragments about 5.0, 2.4, 2.15, and 2.0kb, respectively, different from Archicitrus. Also the pattern of total mtDNAs fragment by the treatment of Pst I showed that the distinguishable fragment pat tern was not appeared in Archicitrus(C. iyo Tanaka), but about 6.0, 5.5, 5.0 and 2.Bkb fragments were appeared only in Metacitrus(C. junos Sieb). Therefore it was indicated that two species in intra-subgenus were identical each other, whereas considerable difference was revealed for inter-subgenus.

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Effects of Rhodobacter sp. SA16 on Lettuce(Lactuca sativa L.) in Plastic Film House (시설 상추에 대한 Rhodobacter sp. SA16 처리 효과)

  • Lee, Young-Han;Jeong, Han-Taek;Yun, Han-Dae
    • Korean Journal of Environmental Agriculture
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    • v.27 no.2
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    • pp.163-170
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    • 2008
  • This experiment was conducted to determine the development of mixed organic fertilizer using photosynthetic bacteria and mass production of mixed microbial compound for the environment-friendly agriculture. Photosynthetic bacteria, Rhodobacter sp. SA16 was isolated from soil collected by plastic film house. The SA16 strain was identified based on 16S rDNA sequence analysis and it is closely related to Rhodobacter sp.(100% similarity). The mixed organic fertilizer using SA16 was made of $N-P_2O_5-K_2O=60-10-20\;g\;kg^{-1}$ with combined soybean cake, sesame cake, powdered blood, fish meal, powdered bones and red-yellow soil. The mixed organic fertilizer 0.45, 0.90 and 1.35 Mg $ha^{-1}$ application in Ihyeon series was treated based on soil testing for lettuce cultivation in plastic film house. These results showed that the yield was increased the 18 and 19%over control by the mixed organic fertilizer application 0.45 and 0.90 Mg $ha^{-1}$, respectively. In the physical properties of the soil, the porosity of mixed organic fertilizer 1.35 Mg $ha^{-1}$ treatment was highest at 58.8%. Our results clearly revealed that the organic fertilizer using Rhodobacter sp. SA16 and mass production of mixed strains could be a useful technology in pursuing environment-friendly agriculture.