• Title/Summary/Keyword: 25S rDNA

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Steroids from the Cold Water Starfish Ctenodiscus crispatus with Cytotoxic and Apoptotic Effects on Human Hepatocellular Carcinoma and Glioblastoma Cells

  • Quang, Tran Hong;Lee, Dong-Sung;Han, Se Jong;Kim, Il Chan;Yim, Joung Han;Kim, Youn-Chul;Oh, Hyuncheol
    • Bulletin of the Korean Chemical Society
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    • v.35 no.8
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    • pp.2335-2341
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    • 2014
  • Chemical investigation on the methanol extract of the starfish Ctenodiscus crispatus resulted in the isolation of five steroids, (22E,$24{\zeta}$)-26,27-bisnor-24-methyl-$5{\alpha}$-cholest-22-en-$3{\beta}$,5,$6{\beta}$,$15{\alpha}$,25-pentol 25-O-sulfate (1), (22E,24R,25R)-24-methyl-$5{\alpha}$-cholest-22-en-$3{\beta}$,5,$6{\beta}$,$15{\alpha}$,25,26-hexol 26-O-sulfate (2), (28R)-24-ethyl-$5{\alpha}$-cholesta-$3{\beta}$,5,$6{\beta}$,8,$15{\alpha}$,28,29-heptaol-24-sulfate (3), (25S)-$5{\alpha}$-cholestane-$3{\beta}$,5,$6{\beta}$,$15{\alpha}$,$16{\beta}$,26-hexaol (4), and ${\Delta}7$-sitosterol (5). Their structures were identified by extensive spectroscopic analyses, including 1D, 2D NMR and MS and chemical methods. Compound 4 showed cytotoxicity against human hepatoma HepG2 and glioblastoma U87MG cells via inhibition of cell growth and induction of apoptosis. Induction of apoptosis by 4 was demonstrated by cell death, DNA fragmentation, increased Bax/Bcl-2 protein ratio and the activation of caspase-3, caspase-9 and poly (ADP-ribose) polymerase (PARP).

Comparative Genomic Analysis of Staphylococcus aureus FORC_001 and S. aureus MRSA252 Reveals the Characteristics of Antibiotic Resistance and Virulence Factors for Human Infection

  • Lim, Sooyeon;Lee, Dong-Hoon;Kwak, Woori;Shin, Hakdong;Ku, Hye-Jin;Lee, Jong-eun;Lee, Gun Eui;Kim, Heebal;Choi, Sang-Ho;Ryu, Sangryeol;Lee, Ju-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.25 no.1
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    • pp.98-108
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    • 2015
  • Staphylococcus aureus is an important foodborne pathogen that causes diverse diseases ranging from minor infections to life-threatening conditions in humans and animals. To further understand its pathogenesis, the genome of the strain S. aureus FORC_001 was isolated from a contaminated food. Its genome consists of 2,886,017 bp double-stranded DNA with a GC content of 32.8%. It is predicted to contain 2,728 open reading frames, 57 tRNAs, and 6 rRNA operons, including 1 additional 5S rRNA gene. Comparative phylogenetic tree analysis of 40 complete S. aureus genome sequences using average nucleotide identity (ANI) revealed that strain FORC_001 belonged to Group I. The closest phylogenetic match was S. aureus MRSA252, according to a whole-genome ANI (99.87%), suggesting that they might share a common ancestor. Comparative genome analysis of FORC_001 and MRSA252 revealed two non-homologous regions: Regions I and II. The presence of various antibiotic resistance genes, including the SCCmec cluster in Region I of MRSA252, suggests that this strain might have acquired the SCCmec cluster to adapt to specific environments containing methicillin. Region II of both genomes contains prophage regions but their DNA sequence identity is very low, suggesting that the prophages might differ. This is the first report of the complete genome sequence of S. aureus isolated from a real foodborne outbreak in South Korea. This report would be helpful to extend our understanding about the genome, general characteristics, and virulence factors of S. aureus for further studies of pathogenesis, rapid detection, and epidemiological investigation in foodborne outbreak.

Flavobacterium jocheonensis sp. nov., Isolated from Marine Green Alga Ulva pertusa

  • Choi, Ha Ri;Park, So Hyun;Heo, Moon Soo
    • Journal of Microbiology and Biotechnology
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    • v.29 no.8
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    • pp.1266-1272
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    • 2019
  • A bacterial strain, labeled $UR11^T$, was isolated from green alga Ulva pertusa collected from Jeju Island, Korea. $UR11^T$ was identified as a gram-negative, rod-shaped, motile by gliding and aerobic bacterial strain with yellow colonies on R2A plates. The strain $UR11^T$ grew over at a temperature range of $10^{\circ}C$ to $30^{\circ}C$ (optimally at $25^{\circ}C$), a pH range of 6.0-11 (optimally at pH 7.0) and a Nacl range of 0.5-5% Nacl (w/v). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain $UR11^T$ was a member of the genus Flavobacterium. Strain $UR11^T$ shared close similarity with F. jejuensis $EC11^T$ (98.0%) F. jumunjinense $HME7102^T$ (96.1%), F. haoranii $LQY-7^T$ (95.3%), F. dongtanense $LW30^T$ (95.1%), and F. ahnfeltiae 10Alg $130^T$(94.9%). The major fatty acids (>5%) were $iso-C_{15:0}$ (33.9%), $iso-C_{15:1}$ G (12.4%), $iso-C_{17:0}$ 3-OH (9.0%), $isoC_{16:0}$ (7.0%) and $iso-C_{15:0}$ 3-OH (6.3%). The major polar lipids were phosphatidylethanolamine, seven unknown aminolipids, two unknown aminopolarlipids and two unknown lipids. DNA-DNA hybridization value was 58% at strain $UR11^T$ with F. jejuensis $EC11^T$. Based on phenotypic, chemotaxonomic and phylogenetic evidence, strain $UR11^T$ represents a novel species of the genus Flavobacterium, for which the name Flavobacterium jocheonensis sp. nov. is proposed. The type strain is Flavobacterium jocheonensis is $UR11^T$ (=KCTC $52377^T$ =JCM $31512^T$).

Sex Determination of Cattle Meat by Polymerase Chain Reaction Amplification of the DEAD Box Protein (DDX3X/DDX3Y) Gene

  • Gokulakrishnan, P.;Kumar, R.R.;Sharma, B.D.;Mendiratta, S.K.;Sharma, D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.5
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    • pp.733-737
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    • 2012
  • Determination of sex origin of cattle meat by fast and reliable molecular methods is an important measure to ensure correct allocation of export refunds particularly in European countries and also female cattle (cow) slaughter is legally banned in India because of religious beliefs. Based on the DEAD box protein gene located on the X and Y chromosomes, 2 pair of primers were designed and the system of PCR was optimized. Upon PCR amplification, male tissue showed 2 bands, while female tissue resulted in only one band. The accuracy and specificity of the primers was assessed using DNA template extracted from cattle meat of known sex. The protocol was subjected to a blind test and showed 100% concordance, proving its accuracy and reliability.

Isolation and Diversity of Yeasts from Fruits and Flowers of Orchard in Sinam-myeon of Yesan-gun, Chungcheongnam-do, Korea (충남 예산군 신암면 일대 과수원의 과일과 꽃들로부터 효모의 분리 및 분포 특성)

  • Hyun, Se-Hee;Lee, Jong-Guk;Park, Won-Jong;Kim, Ha-Kun;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.42 no.1
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    • pp.21-27
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    • 2014
  • Several kinds of yeasts from fruits and flowers of orchard in Yesan-gun of Chungcheongnam-do, Korea were isolated and identified by comparison of nucleotide sequences for PCR-amplified D1/D2 region of 26S rDNA using BLAST. Fourty eight yeast strains of twenty five species and one hundred eight yeast strains of fourty eight species were isolated from fruits and flowers of orchard in Sinam-myeon of Yesan-gun, Chungcheongnam-do, respectively. Among total one hundred fifty-six yeast strains, only sixteen species were overlapped from fruits and flowers.

Isolation and Characterization of Tartaric Acid-Degrading Bacteria from Korean Grape Wine Pomace (국산 포도주 주박으로부터 주석산 분해 세균의 분리 및 특성)

  • Kim, Jong-Hyun;Choi, Sang-Hoon;Hong, Young-A;Kim, Dong-Hwan;Lee, Won-Hee;Rhee, Chang-Ho;Park, Heui-Dong
    • Food Science and Preservation
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    • v.15 no.3
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    • pp.483-490
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    • 2008
  • Several tartaric acid-degrading bacteria were isolated from Korean grape wine pomace after enrichment culture at $30^{\circ}C$ for 10 days in liquid media containing tartaric acid Among them, strains KMBL 5777 and KMBL 5778 exhibited the highest level in the growth and tartaric acid degradability in a medium containing 0.2%(w/v) tartaric acid as a sole carbon source. They were identified as Acetobacter tropicalis based on their morphological and physiological characteristics as well as their 16S rDNA sequences. Blast search of the 16S rDNA sequences revealed that the isolated strains are closest to Acetobacter tropicalis. Homologies of the sequences of KMBL 5777 and KMBL 5778 were 96.0 and 98.9%, respectively with those of A. tropicalis LMG 1663. Both the two bacteria showed higher tartaric acid degradation at $25^{\circ}C$ that those at 20 and $30^{\circ}C$. They could degrade tartaric acid at a wide range of pH between 4.0 and 7.0 with the most rapid degradability at pH 7.0. However, when the bacteria were grown for 8 days, the same level of tartaric acid degradation was observed at pH 4.0, 5.0, 6.0 and 7.0, which was 90.0% of degradation of the acid.

Anti-fungal materials Produced by Streptomyces albogriseus Isolated in Korean soil

  • Kwon, Hyuk-Ku;Kang, Byeong-Kon;Lee, Jang-Hoon
    • Proceedings of the Korean Environmental Health Society Conference
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    • 2004.06a
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    • pp.165-168
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    • 2004
  • An anti-fungal material producing actinomycete was isolated from domestic soil. This strain was identified as Streptomyces albogriseus by 16S rDNA sequence. YEME (yeast extract 4g, malt extract 10g, glucose 4g, D.W 1l , pH $7.0{\pm}0.2$) medium was used for production of anti-fungal materials. S. albogriseus was cultured in a shaking incubator for 2 weeks at 150 rpm and $25{\pm}1^{\circ}C$. An anti-fungal material produced by S. albogriseus was identified at 340nm by uv/vis- spectrometer and it showed powerful anti-fungal activity. This is the first report that secondary metabolite produced by S. albogriseus showed an activity against phytopathogenic fungi such as Collectrichum coccodes, Botrytis cinerea, Cladosporium cucumerinum, Didymella bryoniae.

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Antioxidant Activity and Characterization of Exiguobacterium sp. SC2-1 Isolated from Sea Water (해양에서 분리한 Exiguobacterium sp. SC2-1의 항산화 활성 및 특성)

  • Kim Man-Chul;Park Guen- Tae;Son Hong-Joo;Choi WooBong;Heo Moon-Soo
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.74-80
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    • 2005
  • For the reseach of the natural marine antioxidant, an antioxidant-producing bacterium was isolated from seawater in Jeju costal area. The isolated strain SC2-1 was Gram-positive, catalase positive, oxidase negative, motile and small rods. The strain utilized sucrose, dextrose, fructose, mannitol and maltose as a sole carbon and energy source and NaCl required for growth. The radical scavenging activity of the culture supernatant was detemined by DPPH method. This bacterium was identified based on morphological, biochemical characteristics and 16S rDNA sequence analysis, and then was named Exiguobacterium sp. SC2-1. The optimum conditions of culture for production antioxidnat were $25^{\circ}C$, pH 6-8 and $4\%$ NaCl. The stain showed the highest activity and growth cultured in medium which added $1\%$ maltose. Hydroxyl radical activity of the supernatant of Exiguobacterium sp. SC2-1 was $73\%$. The SOD activity of the culture supernatant was estimated about $35\%$.

Isolation of Tetracycline-resistant Lactic Acid Bacteria from Kimchi (김치에서 tetracycline 내성 유산균의 분리)

  • Kang, Hyo-Jin;Kim, Byung-Chun;Park, Wan
    • Korean Journal of Microbiology
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    • v.40 no.1
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    • pp.1-6
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    • 2004
  • Tetracycline resistant bacterial strains were isolated from 10 batches of Kimchi among 50 batches collected in Taegu restrict. The MIC of tetracycline ranged between 25 and> 100 ㎖/l. Total genomic DNA preparation from all 10 tetracycline resistant lactic acid bacterial isolates were subjected to PCR amplification with class-specific primers for tet(M) and tet(O). In only one isolate, HJ9, tet(M) was detected. By Southern blotting and hybridization with a tet(M)-specific probe, the tet(M) gene of HJ9 isolate could be localized on a plasmid. The partial nucleotide sequence and deduced amino acid sequence of tet(M) of HJ9 showed 90-99% and 94-100% homology to those of Gram positive bacteria, respectively. With sequencing of 16S rRNA, HJ9 isolate from Kimchi was identified as Lactobacillus sakei. From these results, Kimchi can be considered potential vehicle for the spread of antibiotic-resistant lactic acid bacteria along the food chain to the consumer.

Isolation and Identification of Wild Yeast and Its Use for the Production of Grapewine (야생 효모의 분리.동정 및 이를 이용한 포도주 제조)

  • Kim, Jung-In;Lee, Nam-Keun;Hahm, Young-Tae
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.217-221
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    • 2007
  • The domestic cultured Campbell's Early and Geubong grapes were fermented far the production of red wines with the isolated wild yeast Saccharomyces cerevisiae IJ850. For the isolation of wild yeast, Geubong and Campbell's Early grapejuices were naturally fermented at room temperature for 6 days without adding stater culture. The strain isolated from Geubong which has 1.8 times higher fermentative ability than the strains isolated Campbell Early was selected. The selected strain was identified by using 26S rDNA sequencing. The strain showed 99.7% of similarity with Saccharomyces cerevisiae and thus identified as Saccharomyces cerevisiae IJ850. It was investigated the fermentative ability as the start culture. For the production of grapewine, the final sugar concentrations of grapejuices were adjusted to the $25^{\circ}Brix$ with anhydrous glucose. The grapejuices were fermented at room temperature for 10 days in the air-locked bottles filled with $CO_2$ gas. The final yield and alcohol concentration of Campbell's Early and Geubong grapewines fermented with the isolated wild yeast were 80.8%, 11.0% and 87.8%, 13.0%, respectively. Between the isolated wild yeast S. cerevisiae IJ850 and the commercial yeast S. cerevisiae EC1118, total acidities of grapewines produced with wild yeast were lower than those produced with the commercial yeast. The pH values and the values of color analysis of grapewines produced with both strains were similar. The total phenol contents of campbell's Early and Geubong wines produced with the isolated yeast and the commercial yeast were obtained in the range of 75 to 125mg/L. In conclusion, S. cerevesiae IJ850 isolated from the domestic cultured Geubong grape is able to use to produce grapewines as stater culture.