• Title/Summary/Keyword: rRNA

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The Phylogenetic Analysis of the NS-5 Region Sequence of Hepatitis G Viruses Isolated in Korea (국내에서 분리된 G형 간염바이러스 NS-5 Region 염기서열의 계통학적 분석)

  • Jee, Young-Mee;Kim, Ki-Soon;Cheon, Doo-Sung;Park, Jeong-Koo;Kang, Young-Hwa;Lee, Yoon-Sung;Chung, Yoon-Suk;Kim, Ji-Eun;Yoon, Jae-Deuk
    • The Journal of Korean Society of Virology
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    • v.29 no.1
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    • pp.45-53
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    • 1999
  • We examined the hepatitis G virus infections among 227 Koreans who were healthy or were suspected of hepatitis and determined the phylogenetic relationship based on a part of the NS-5 region of 5 positive samples. Viral RNA was extracted from sera and cDNA was synthesized and subsequently amplified by RT-PCR (reverse transcription-polymerase chain reaction) or RT-nested PCR using random hexamer and NS-5 specific primers (470-20-1-77F, 470-20-1-211R, HGVNESTFO, HGVNESTRE). Five positives were found to belong to samples of patients showing symptoms of viral hepatitis. Primers used for PCR or nested PCR were derived from the NS-5 region. On the other hand, no amplification was detected using primers derived from the 5'-NCR (G-146F, G-401R). We performed TA cloning and sequencing of 5 amplified fragments, and their sequences were compared with those of foreign isolates of HGV. The phylogenetic analysis using MegAlign programme of DNAstar has shown that the Korean isolates are clustered on the phylogenetic tree. In summary, we confirmed the hepatitis G virus infection in 5 cases out of 12 patients showing the symptoms of viral hepatitis. The phylogenetic analysis of sequences of 5 amplified fragments showed that their relations to each other were closer than those to the foreign HGV isolates reported.

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Production and Characteristics of Lytic Enzyme against Streptococcus mutans Cell Wall from Alkalophilic Bacillus sp. 4830 (호알카리성 Bacillus sp. 4830이 생산하는 Streptococcus mutans 세포벽 분해효소의 분리와 특성)

  • Kim, Yun-Keun;Bai, Dong-Hoon
    • Korean Journal of Food Science and Technology
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    • v.35 no.6
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    • pp.1143-1149
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    • 2003
  • To elucidate a method of preventing dental caries, strains producing lytic enzymes were isolated and their characteristics were investigated. Among 5,00 alkalophilic strains isolated from soil, 22 strains showed lytic activity against Streptococcus mutans. Strain No. 4830, with the highest lytic activity, was selected for further study. Strain 4830 showed 94% sequence homology with the 16S rDNA sequence of Bacillus alcalophilus, but it was concluded to be different from Bacillus alcalophilus because of its biochemical characteristics. The strain was named Bacillus sp. 4830. The lytic enzyme from Bacillus sp. 4830 was purified by ethanol precipitation and CM-agarose column chromatography. The molecular weight of the lytic enzyme was determined to be 28 kDa by SDS-PAGE. The lytic enzyme was stable between pH 5.0 and pH 11 and up to $40^{\circ}C$. The optimal pH and temperature for the lytic activity was 9.0 and $50^{\circ}C$, respectively.

Optimization of Indole-3-Acetic production by phosphate solubilization bacteria isolated from waste mushroom bed of Agaricus bisporus

  • Walpola, Buddhi Charana;Noh, Jae-Geun;Kim, Chan Kyem;Kyung, Ki-Cheon;Kong, Won-Sik;Yoon, Min-Ho
    • Journal of Mushroom
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    • v.11 no.2
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    • pp.53-62
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    • 2013
  • A total of 35 phosphate solubilizing bacterial strains were isolated from waste mushroom bed of Agaricus bisporus in Buyeo-Gun, Chungnam and screened for the production of indole acetic acid (IAA). The best IAA producing strain was identified as Pantoea rodasii using 16S rRNA analysis. In addition to the IAA production, this strain could act as an efficient phosphate solubilizer (1100 ${\mu}g$ $ml^{-1}$ after 5 days of incubation) also. The selected strain was cultured under different conditions in order to assess the optimum conditions for maximum IAA production. The nutrient broth (NB) medium was recorded as the best medium, where the maximum IAA production (229 ${\mu}g$ $ml^{-1}$) was recorded at the start of stationary phase (12 hours after inoculation) of the bacteria growth. The performance of the strain was found to be maximum at the temperature of $30^{\circ}C$ followed by $25^{\circ}C$. IAA production was found to be increased with increasing tryptophan concentration (from 0.1 to 0.6%), however beyond this limit, a slight reduction in IAA production was observed. The strains' ability to produce IAA was further confirmed by extraction of crude IAA and subsequent TLC analysis. A specific spot from the extracted IAA preparation was found corresponding with the standard spot of IAA with same $R_f$ value. The results of HPLC analysis conducted in identifying and quantifying the IAA production more precisely, are in agreement with the results of the assessment done with colorimetric method. As revealed by the results of the pot experiment, the isolated strain could significantly enhance the growth (as measured by shoot and root growth) of mung bean plants compared to that of non-inoculated plants. Therefore it can be concluded that the present strain, Pantoea rodasii has great potential to be used as bio-inoculants.

VP7 Genotypes of Group A Rotavirus Isolated from Infants and Toddlers with Rotavirus Gastroenteritis in Jeju (제주지역 로타바이러스 위장관염 환아로부터 분리한 A군 로타바이러스의 VP7 Genotypes에 대한 연구)

  • Kang, Ki Soo;Shin, Kyung-Sue;Cui, Xiu Ji;Kim, Wonyong
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.9 no.2
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    • pp.147-152
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    • 2006
  • Purpose: Efficacy of the new rotavirus vaccines ($Rotarix^{(R)}$, $RotaTeq^{(R)}$) recently developed can be affected by the rotavirus genotypes prevalent in communities. We performed this study to identify the recent distribution of rotavirus genotypes prevalent in Jeju. Methods: Genotyping of human rotaviruses was performed using 81 samples collected from 154 inpatients and outpatients with rotavirus gastroenteritis at Cheju National University Hospital between July 2005 and June 2006. All six (1, 2, 3, 4, 8, 9) G serotypes were identified by amplification of segments of the gene for VP7 using the reverse transcription-polymerase reaction (RT-PCR). Results: The results of RT-PCR for 81 samples were all positive. G typing of the VP7 protein showed that G1 was the most dominant circulating genotype (65.5%) followed by G2 (14.8%), G3 (13.6%), G8 (1.2%), G9 (1.2%), G4 (0%), and a combination of G1/G3 (3.7%). Conclusion: This distribution of rotavirus VP7 genotypes in Jeju is different from that in other domestic areas; the most dominant circulating genotype was G1.

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Physiological Characteristics and ACE Inhibitory Activity of Lactobacillus zeae RMK354 Isolated from Raw Milk (원유에서 분리한 Lactobacillus zeae RMK354의 생리적 특성 및 ACE 억제능)

  • Lim, Sang-Dong;Kim, Kee-Sung;Do, Jeong-Ryong
    • Food Science of Animal Resources
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    • v.28 no.5
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    • pp.587-595
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    • 2008
  • In order to develop a new starter for fermented milk, 1037 bacterial strains were isolated from raw milk. The strain that showed excellent acid producing and angiotensin converting enzyme (ACE) inhibitory activity (88.6%) was selected and identified as a Lactobacillus zeae based on the result of API carbohydrate fermentation pattern and 16S rDNA sequence. Lactobacillus zeae RMK354 was investigated further to study its physiological characteristics. It showed strong ACE inhibitory activity compared with commercial LAB starters tested. The optimum growth temperature of L. zeae RMK354 was $40^{\circ}C$ and it took 10 hr to reach pH 4.3 under this condition. L. zeae RMK354 showed more sensitive to penicillin-G, bacitracin, novobiocin, in a comparison of 14 different antibiotics, and showed most resistance to polymyxin B and vancomycin. It showed higher esterase and leucine arylamidase activities compared with 16 other enzymes. It was comparatively tolerant to bile juice and able to survive at pH 2 for 3 hr. It showed inhibitory activity against Salmonella Typhimurium with the rate of 60%. Based on these and previous results, L. zeae RMK354 could be an excellent starter culture for fermented milk with high level of ACE inhibitory activity.

Culture Conditions for Improving Extracellular Lipolytic Enzyme Production by a Novel Thermophilic Geobacillus sp. AR1 (신규 고온성 Geobacillus sp. AR1의 extracellular 지질분해효소 생산을 위한 배양조건)

  • Park, Su-Jin;Jeon, Sung-Jong
    • Journal of Life Science
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    • v.23 no.1
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    • pp.110-115
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    • 2013
  • A microorganism (strain AR1) producing an extracellular lipolytic enzyme was isolated from hot springs located in Beppu, Japan. Phylogenetic analysis based on the 16S rDNA sequence and biochemical studies indicated that AR1 belongs to the genus Geobacillus. This study focused on novel strategies to increase extracellular lipolytic enzyme production by this novel Geobacillus sp. AR1. Cultures of the AR1 strain grew within a wide temperature range (from 35 to $75^{\circ}C$); the optimum temperature was $65^{\circ}C$. The pH for optimal growth was 6.5, whereas the optimum pH for lipolytic enzyme production was 8.5. The presence of oils in the culture medium led to improvements in lipolytic enzyme activity. Soybean oil was the most efficient inducer, and it yielded better results when added in the exponential phase. On the other hand, the addition of chemical surfactants led to lipolytic enzyme production. Their addition to the culture could affect the location of the enzyme activity. The addition of Tween 20 in the stationary phase significantly increased the proportion of the extracellular enzyme activity. According to the results, following the addition of soybean oil and Tween 20 in the exponential and stationary phases, the extracellular lipolytic activity was increased 2.4-fold compared with that of a control.

Antifungal activity against cheese fungi by lactic acid bacteria isolated from kimchi (김치 분리 유산균의 치즈 곰팡이 항진균 활성)

  • Choi, Ha Nuel;Oh, Hyun Hee;Yang, Hee Sun;Huh, Chang Ki;Bae, In Hyu;Lee, Jai Sung;Jeong, Yong Seob;Jeong, Eun Jeong;Jung, Hoo Kil
    • Food Science and Preservation
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    • v.20 no.5
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    • pp.727-734
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    • 2013
  • The antifungal activity against cheese fungi by lactic acid bacteria isolated from kimchi was investigated. Eight fungi were isolated from cheese in the cheese ripening room. Two of them were identified as Penicillium and Cladosporium via ITS-5.8S rDNA analysis. Twenty-two species of lactic acid bacteria with antifungal activity were isolated from kimchi. Two of them were identified as Lactobacillus and Pediococcus via 16S rRNA sequence analysis. Of the 22 lactic acid bacteria species, six were selected (L. sakei subsp. ALJ011, L. sakei subsp. ALI033, L. sakei subsp. ALGy039, P. pentosaceus ALJ015, P. pentosaceus ALJ024 and P. pentosaceus ALJ026) due to their higher activity against the eight fungi isolated from cheese in the cheese ripening room; and among the six species, the P. pentosaceus ALJ015 and P. pentosaceus ALJ024 isolates from the Jeonju area kimchi and the L. sakei subsp. ALI033 isolate from the Iimsil area kimchi had higher antifungal activity than the other lactic acid bacteria. The minimum inhibitory concentration (MIC) of L. sakei subsp. ALI033 against the eight fungi isolated from cheese in the cheese ripening room was $62.5{\mu}g/mL$.

Korean Red Ginseng attenuates anxiety-like behavior during ethanol withdrawal in rats

  • Zhao, ZhengLin;Kim, Young Woo;Wu, YiYan;Zhang, Jie;Lee, Ju-Hee;Li, XiaoHua;Cho, Il Je;Park, Sang Mi;Jung, Dae Hwa;Yang, Chae Ha;Kim, Sang Chan;Zhao, RongJie
    • Journal of Ginseng Research
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    • v.38 no.4
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    • pp.256-263
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    • 2014
  • Background: Korean Red Ginseng (KRG) is known to have antianxiety properties. This study was conducted to investigate the anxiolytic effects of KRG extract (KRGE) during ethanol withdrawal (EW) and the involvement of the mesoamygdaloid dopamine (DA) system in it. Methods: Rats were treated with 3 g/kg/d of ethanol for 28 d, and subjected to 3 d of withdrawal. During EW, KRGE (20 mg/kg/d or 60 mg/kg/d, p.o.) was given to rats once/d for 3 d. Thirty min after the final dose of KRGE, anxiety-like behavior was evaluated in an elevated plus maze (EPM), and plasma corticosterone (CORT) levels were determined by a radioimmunoassay (RIA). In addition, concentrations of DA and 3,4-dihydroxyphenylacetic acid (DOPAC) in the central nucleus of the amygdala (CeA) were also measured by high performance liquid chromatography (HPLC). Results: The EPM test and RIA revealed KRGE inhibited anxiety-like behavior and the over secretion of plasma CORT during EW. Furthermore, the behavioral effect was blocked by a selective DA D2 receptor (D2R) antagonist (eticlopride) but not by a selective DA D1 receptor (D1R) antagonist (SCH23390). HPLC analyses showed KRGE reversed EW-induced decreases of DA and DOPAC in a dose-dependent way. Additionally, Western blotting and real-time polymerase chain reaction (PCR) assays showed that KRGE prevented the EW-induced reductions in tyrosine hydroxylase (TH) protein expression in the CeA and TH mRNA expression in the ventral tegmental area (VTA). Conclusion: These results suggest that KRGE has anxiolytic effects during EW by improving the mesoamygdaloid DA system.

Characteristics of yeast with low temperature adaptation for Yakju brewed (약주 제조를 위한 저온 적응성 효모의 특성)

  • Seo, Dong-Jun;Yeo, Soo-Hwan;Mun, Ji-Young;Jung, Woo-Jin;Cho, Yong Sik;Baek, Seong Yeol
    • Food Science and Preservation
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    • v.22 no.6
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    • pp.908-914
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    • 2015
  • The objectives of this study were to isolate and characterize low temperature adaptation yeast and to obtain suitable yeasts strains for manufacturing Yakju. In this study, we isolated 482 wild yeasts from fermented foods. Out of these, 5 yeast strains were selected based on increased growth at low temperature ($15^{\circ}C$) and high ${\beta}$-glucosidase activity. To screen the aromatic level of isolates, media containing cerulenin and 5,5,5-trifluor-DL-leucine (TFL) were used. Y297 strain demonstrated tolerance against TFL and produced more than 13% alcohol. Y297 strain was identified a Saccharomyces cerevisiae based on the 26S rDNA gene sequences. Maximum cell growth was observed after 19 hr and 38 hr of incubation at $25^{\circ}C$ and $15^{\circ}C$, respectively. The exponential phase was followed by a lengthy stationary phase, at $15^{\circ}C$, when the cells remained high viable. Y297 strain demonstrated tolerance against alcohol (10%), glucose (60%) and salt(NaCl, 8%). ${\beta}$-glucosidase and esterase activity in Y297 were higher than those of controls at $15^{\circ}C$. Overall, these results indicated that using wild yeast strain, isolated from fermented food, affects the chemical characteristics of the brewed Yakju.

Isolation of Mannanase-producing Bacteria, Bacillus subtilis WL-6 and WL-11, and Cloning and Characterization of Mannanase (Bacillus subtilis 분리균 2주 유래 mannanases의 특성 비교)

  • Yoon, Ki-Hong
    • Journal of Life Science
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    • v.26 no.10
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    • pp.1113-1120
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    • 2016
  • Two bacterial strains producing extracellular man nanase were isolated from doenjang, a traditionally fermented soybean paste in Korea. The isolates, WL-6 and WL-11, were identified as Bacillus subtiis on the basis of their 16S rRNA gene sequences, morphological, and biochemical properties. Two genes encoding the mannanase of both B. subtilis WL-6 and B. subtilis WL-11 were each cloned into Escherichia coli, and their nucleotide sequences were determined. Both mannanase genes consisted of 1,086 nucleotides, encoding polypeptides of 362 amino acid residues. The deduced amino acid sequences of the two WL-6 and WL-11 mannanases, designated Man6 and Man11, respectively, differed from each other by eight amino acid residues, and they were highly homologous to those of mannanases belonging to the glycosyl hydrolase family 26. The 26 amino acid stretch in the N-terminus of Man6 and Man11 was a predicted signal peptide. Both Man6 and Man11 were localized at the level of 94–95% in an intracellular fraction of recombinant E. coli cells. The enzymes hydrolyzed both locust bean gum and mannooligosaccharides, including mannotriose, mannotetraose, mannopentaose, and mannohexaose, forming mannobiose and mannotriose as predominant products. The optimal reaction conditions were 55°C and pH 6.0 for Man6, and 60°C and pH 5.5 for Man11. Man11 was more stable than Man6 at high temperatures.