• Title/Summary/Keyword: ITS rDNA

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The interaction of serum albumin with ginsenoside Rh2 resulted in the downregulation of ginsenoside Rh2 cytotoxicity

  • Lin, Yingjia;Li, Yang;Song, Zhi-Guang;Zhu, Hongyan;Jin, Ying-Hua
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.330-338
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    • 2017
  • Background: Ginsenoside Rh2 (G-Rh2) is a ginseng saponin that is widely investigated because of its remarkable antitumor activity. However, the molecular mechanism by which (20S) G-Rh2 triggers its functions and how target animals avoid its cytotoxic action remains largely unknown. Methods: Phage display was used to screen the human targets of (20S) G-Rh2. Fluorescence spectroscopy and UV-visible absorption spectroscopy were used to confirm the interaction of candidate target proteins and (20S) G-Rh2. Molecular docking was utilized to calculate the estimated free energy of binding and to structurally visualize their interactions. MTT assay and immunoblotting were used to assess whether human serum albumin (HSA), bovine serum albumin (BSA), and bovine serum can reduce the cytotoxic activity of (20S) G-Rh2 in HepG2 cells. Results: In phage display, (20S) G-Rh2-beads and (20R) G-Rh2-beads were combined with numerous kinds of phages, and a total of 111 different human complementary DNAs (cDNA) were identified, including HSA which had the highest rate. The binding constant and number of binding site in the interaction between (20S)-Rh2 and HSA were $3.5{\times}10^5M^{-1}$ and 1, and those in the interaction between (20S) G-Rh2 and BSA were $1.4{\times}10^5M^{-1}$ and 1. The quenching mechanism is static quenching. HSA, BSA and bovine serum significantly reduced the proapoptotic effect of (20S) G-Rh2. Conclusion: HSA and BSA interact with (20S) G-Rh2. Serum inhibited the activity of (20S) G-Rh2 mainly due to the interaction between (20S) G-Rh2 and serum albumin (SA). This study proposes that HSA may enhance (20S) G-Rh2 water solubility, and thus might be used as nanoparticles in the (20S) G-Rh2 delivery process.

Identification of Microorganisms, Cladosporium sp. and Sterigmatomyces sp., Proliferated on the Surface of Traditional Soy Sauce, and the Effect of NaCl Concentration on Their Enzymatic Activity (전통간장으로부터 분리한 Cladosporium sp.과 Sterigmatomyces sp. 미생물의 동정 및 NaCl 농도에 따른 미생물 효소활성 분석)

  • Lee, Nam-Keun;Ryu, Young-Jun;Yeo, In-Cheol;Park, Sung-Joon;Kwon, Ki-Ok;Cha, Chang-Jun;Hahm, Young-Tae
    • Korean Journal of Food Science and Technology
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    • v.44 no.4
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    • pp.488-492
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    • 2012
  • Two strains, traditionally referred to as rock flower (Bawhi-kot) and buckwheat flower (Memil-kot or Chile-Kot), were isolated from stored traditional soy sauce and were identified by using the 18S ITS1/4 region sequences. The rock flower strain showed 99% of similarity with Cladosporium sp. and buckwheat flower strain was 99% identical with yeast Sterigmatomyces halophilus. Both strains were tentatively named Cladosporium sp. NK1 and Sterigmatomyces halophilus NK2, respectively. The optimal growth pHs and temperatures of both strains in a YPD broth medium were in the range of pH 5.0 to 7.0 and 22 to $27^{\circ}C$. Both strains were able to grow in more than 20% of NaCl. In the enzyme activity assay, high protease activity of Cladosporium sp. NK1 and S. halophilus NK2 were obtained in YPD containing 10% of NaCl. High amylase activities of both stains were in 15% and 5% of NaCl, respectively. Lipase activity was, however, not detected in both strains.

Production and Characterization of Alkaline Protease of Micrococcus sp. PS-1 Isolated from Seawater (해수에서 분리한 Micrococcus sp. PS-1이 생산하는 단백질 분해효소의 생산과 효소학적 특성)

  • Jin, Young-Rang;Yu, Sun-Nyoung;Kim, Kwang-Youn;Kim, Sang-Hun;Park, Seul-Ki;Kim, Hyeun-Kyeung;Lee, Yong-Seok;Choi, Yong-Lark;Ji, Jae Hoon;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.23 no.2
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    • pp.273-281
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    • 2013
  • The purpose of this research was to investigate the production and characterization of alkaline protease from Micrococcus sp. PS-1 newly isolated from seawater. Micrococcus sp. PS-1 was grown in Luria-Bertani (LB) medium. Its optimal temperature and pH for growth were $30^{\circ}C$ and 7.0, respectively. The effect of nitrogen sources was investigated on optimal enzyme production. A high level of alkaline protease production occurred in LB broth containing 2% skimmed milk. The protease was purified in a 3-step procedure involving ultrafiltration, acetone precipitation, and dialysis. The procedure yielded a 16.43-purification fold, with a yield of 54.25%. SDS-PAGE showed that the enzyme had molecular weights of 35.0 and 37.5 kDa. Its maximum protease activity was exhibited at pH 9.0 and $37^{\circ}C$, and its activity was stable at pH 8.0-11.0 and $25-37^{\circ}C$. The protease activity was strongly inhibited by PMSF, EDTA, and EGTA. Taken together, the results demonstrate that the protease enzyme from Micrococcus sp. PS-1 probably belongs to a subclass of alkaline metallo-serine proteases.

Spatial Distribution and Host Plants of the Ramulus koreanus (Phasmida; Phasmatidae) in Korean Thistle Cultivation (고려엉겅퀴 재배지에서 발생한 우리대벌레 공간분포 및 기주식물)

  • Son, Minwoong;Jung, Chung Ryul;Kwon, Gimyon;Jung, Chuleui
    • Korean journal of applied entomology
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    • v.59 no.4
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    • pp.281-293
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    • 2020
  • The Korean thistle (Cirsium setidens) is grown in Gangwon province as a food ingredient called gondre. Based on regular monitoring of Korean thistle cultivation, we detected local outbreaks of a stick insect causing severe damage. Here we report the identification of the stick insect and its ecological characteristics. The survey involved 3 farms located in Jeongeon-gun, Gangwon province and lasted from May 28 to October 1 in 2019. According to morphological and DNA analyses the stick insect was identified as Ramulus koreanus Kwon Ha and Lee. a member of the family Phasmatidae. Its outbreak in the Korean thistle farms was observed from June 11 to August 22 with peak density on July 23. Spatial distribution analyses, using Taylors power law and Greens index showed that R. koreanus was uniformally distributed. Food consumption rate of Korean thistle leaves was estimated as 60.98±4.35 ㎠/day/adult. The main food plants were identified as Juglans mandshurica and Aronia melanocarpa. Further study needs to pursue the mechanism of the outbreak and its possible impact on agricultural and forest ecosystems. Additionally, means to relieve crop damage need to be identified.

Antimicrobial Drug Resistance and R-plasmid of Salmonella species (Salmonella 균속의 항균제 내성 및 R-plasmid)

  • Lee Myung-Won;Chung Tae-Wha;Lee Yun-Tai;Kang Jeung-bok
    • Journal of environmental and Sanitary engineering
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    • v.3 no.2 s.5
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    • pp.23-41
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    • 1988
  • Two hundred and eighty-six strains of Salmonella species were isolated from the twelve provincial institutes of health and 19 general hospitals of urban and rural areas in Korea from January to December in 1986. The antimicrobial susceptibility test of these cultures was done by the method of agar diluton. The resistance frequency of Salmonella cultures was $29.7\%$. Among these resistant cultures, the most provalent resistance pattern of Salmonella was ampicillin, carbenicillin, chloramphenicol, tetracycline, streptomycin, and its resistance frequency was $15\%$. In plasmid profile of resistance strains, average number of plasmid harboring in Salmonella was 1-4 and molecular weight of plasmid ranged 1.6 to 70 megadalton (Md.). Plasmid pattern of strains isolated from Seoul and Kang-won showed the same or similar profiles. Plasmid pattern was identical in the same resistance pattern.

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THE ANTICANCER EFFECT OF PACLITAXEL($Taxol^{(R)}$) IN ORAL SQUAMOUS CELL CARCINOMA XENOGRAFT (이종 이식된 구강편평세포 암종에서 Paclitaxel ($Taxol^{(R)}$)의 항암 효과)

  • Kim, Ki-Hwan;Kim, Chul-Hwan;Han, Se-Jin;Lee, Jae-Hoon
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.2
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    • pp.95-110
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    • 2006
  • The treatment for oral and maxillofacial carcinoma with chemotherapeutic agents is evaluated by many effective methods to reduce the tumor mass and cancer cell proliferation. However these chemotherapy have many serious side effects, such as bone marrow suppression, renal toxicity, G-I troubles. Therefore a possible approach to develop a clinically applicable chemotherapeutic agent is to screen anticancer activity of Taxol which is known to have very little side effect and have been used to breast cancer and ovarian carcinoma. Taxol is a new anti-microtubular anti-cancer agent extracted from the bark of the Pacific yew, Taxus brevifolia. Paclitaxel(Taxol) acts by promoting tubulin polymerization and over stabilizing microtubules agianst depolymerization. Despite the constant improvements of methods of the cancer treatment especially chemotherapy, the rate of cancer metastasis and recurrent are not decreased. Thus the investigation of new drug which have very little side effect and a possible clinically application continues to be a high priority. Considering that the Taxol have shown very effective chemotherapeutic agent with relatively low toxicity in many solid tumors, it deserves to evaluate its efficacy in oral squamous cell carcinoma. In this study, to investigate the in-vivo and in-vitro anti-cancer efficacy of Taxol in oral squamous cell carcinoma and lastly, the potency of Paclitaxel in the clinical application for oral cancer was evaluated. In vivo study, after HN22 cell line were xenografted in nude mice, the growth of tumor mass was observed, 3 mg/Kg taxol was injected intraperitoneally into nude mice containing tumor mass. The methods of these study were measurement of total volume of tumor mass, histopathologic study, immunohistochemical study, drug resistance assay, growth curve, MTT assay, flow cytometry, cDNA microarray in vivo and in vitro. The results were obtained as following. 1. The visual inspection of the experimental group showed that the volume of the tumor mass was slightly decreased but no significant difference with control group. 2. Ki-67 index was decreased at weeks 4 in experimental group. 3. Microscopic view of the xenografted tumor mass showed well differentiated squamous cell carcinoma and after Taxol injection, some necrotic tissue was seen weeks 4. 4. The growth curve of the tumor cells were decreased after 1day Taxol treatment. 5. According to the MTT assay, HN22 cell line showed relative drug resistancy above $5\;{\mu}g/ml$ concentrations of Taxol. 6. In drug resistance assay, the decrease of cell counts was seen relatively according to concentration. 7. In Flow cytometry, G2M phase cell arrests were seen in low concentration of the Taxol, while S phase cell arrests were seen in high concentration of the Taxol. 8. Using cDNA microarray technique, variable gene expression of ANGPTL4, TXNRD1, FAS, RRAGA, CTGF, CYCLINEA, P19, DUSP5, CEBPG, BTG1 were detacted in the oral squamous cell carcinoma cell after taxol treatment. In this study paclitaxel is effective against oral squamous cell carcinoma cell lines in vitro, but week effect was observed in vivo. So we need continuous study about anticancer effect of taxol in vivo in oral squamous cell carcinoma.

Identification and Sequence Analysis of RNA3 of a Resistance-Breaking Cucumber mosaic virus Isolate on Capsicum annuum

  • Lee Mi-Yeon;Lee Jang-Ha;Ahn Hong-Il;Yoon Ju-Yeon;Her Nam-Han;Choi Jang-Kyung;Choi Gug-Seon;Kim Do-Sun;Harn Chee-Hark;Ryu Ki-Hyun
    • The Plant Pathology Journal
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    • v.22 no.3
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    • pp.265-270
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    • 2006
  • Cultivated hot pepper crops showing severe mosaic symptom were found in Korea in 2004 and their causal agent was identified as Cucumber mosaic virus (CMV). These pepper crops was resistant to the virus in the filled, and they belonged to pathotype 0 (P0) resistant pepper. Resistance screening of selected pepper plants showed that a pepper isolate of CMV was the P0 resistance-breaking virus. This P0 resistance-breaking isolate of CMV, named as Ca-P1, was isolated from leaves of the virus-infected Capsicum annuum cv. Manidda that showed systemic severe mosaic symptom. Ca-P1-CMV could induce systemic mosaic symptoms on P0-susceptible (P0-S) and P0-resistant (P0-R) cultivars whereas an ordinary strain (Fny-CMV) could not infect P0-R. This result suggests that Ca-P1-CMV can overcome P0 resistant pepper cultivars. To analyze its genome sequence, the complete nucleotide sequence of RNA3 of Ca-P1-CMV was determined from the infectious full-length cDNA clone of the virus. RNA3 of Ca-P1-CMV consisted of 2,219 nucleotides. Overall sequence homology of RNA3-encoded two viral proteins (movement protein and coat protein) revealed high similarity (75.2-97.2%) with the known CMV strains. By sequence analysis with known representative strains of CMV, Ca-P1-CMV belongs to a typical member of CMV subgroup IB. The resistance and resistance-breaking mechanisms of pepper and counterpart CMV, respectively, remain to be investigated, which will enrich the genetic resources and accelerate CMV-resistant pepper breeding programs.

Molecular Genetic Identification of Yeast Strains Isolated from Egyptian Soils for Solubilization of Inorganic Phosphates and Growth Promotion of Corn Plants

  • Hesham, Abd El-Latif;Mohamed, Hashem M.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.1
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    • pp.55-61
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    • 2011
  • Forty yeast strains isolated from soils taken from different locations in Egypt were tested for their P-solubilizing activities on the basis of analyzing the clear zone around colonies growing on a tricalcium phosphate medium after incubation for 5 days at $25^{\circ}C$, denoted as the solubilization index (SI). Nine isolates that exhibited P-solubilization potential with an SI ranging from 1.19 to 2.76 were genetically characterized as five yeasts belonging to the genus Saccharomyces cerevisiae and four non-Saccharomyces, based on a PCR analysis of the ITS1-26S region amplied by SC1/SC2 species-specific primers. The highest P-solubilization efficiency was demonstrated by isolate PSY- 4, which was identified as Saccharomyces cerevisiae by a sequence analysis of the variable D1/D2 domain of the 26S rDNA. The effects of single and mixed inoculations with yeast PSY-4 and Bacillus polymyxa on the P-uptake and growth of corn were tested in a greenhouse experiment using different levels of a phosphorus chemical fertilizer (50, 100, and 200 kg/ha super phosphate 15.5% $P_2O_5$). The results showed that inoculating the corn with yeast PSY-4 or B. polymyxa caused significant increases in the shoot and root dry weights and P-uptake in the shoots and roots. The P-fertilization level also had a significant influence on the shoot and root dry weights and P-uptake in the shoots and roots when increasing the P-level from 50 up to 200 kg/ha. Dual inoculation with yeast strain PSY-4 and B. polymyxa at a P-fertilization level of 200 kg/ha gave higher values for the shoot and root dry weights and P-uptake in the shoots and roots, yet these increases were nonsignificant when compared with dual inoculation with yeast strain PSY-4 and B. polymyxa at a P-fertilization level of 100 kg/ha. The best increases were obtained from dual inoculation with yeast strain PSY-4 and B. polymyxa at a P-fertilization level of 100 kg/ha, which induced the following percentage increases in the shoot and root dry weights, and P-uptake in the shoots and roots; 16.22%, 46.92%, 10.09%, and 31.07%, respectively, when compared with the uninoculated control (fertilized with 100 kg/ha).

Rhizopus Soft Rot on Lily Caused by Rhizopus oryzae in Korea (Rhizopus oryzae에 의한 백합 무름병)

  • Hahm, Soo-Sang;Hong, Gye-Wan;Kim, Byung-Ryun;Han, Kwangseop;Choi, Takyong;Nam, Yungyu;Yu, Seunghun
    • Research in Plant Disease
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    • v.20 no.1
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    • pp.50-53
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    • 2014
  • Rhizopus soft rot of lily (Lilium longiflorum) caused by Rhizopus oryzae was observed in the experimental field in Taean Lily Experiment Station in Korea, 2012. The typical symptoms were water-soaked lesions on bottom stem and leaf rot. The lesion rapidly expanded and the plant was softened totally. The fungus grew vigorously at an optimum temperature ($25^{\circ}C$) and brownish colony and black sporangia were formed on potato dextrose agar medium. Sporangiophores formed on end of sporangia were sub-globose, brownish and $6-10{\mu}m$ in size. Sporangia were globose, blackish and $87-116{\mu}m$ in size. Sporangiospores were irregularly oval and sub-globose, brownish $4-8{\mu}m$ in size. On the basis of mycological characteristics, analyzing sequences of internal transcribed spacer region of ribosomal DNA, and pathogenicity test on host plants, the causal fungus was identified as R. oryzae. This is the first report of Rhizopus soft rot on lily caused by R. oryzae in Korea.

Characterization of Cellulase and Xylanase from Bacillus subtilis NC1 Isolated from Environmental Soil and Determination of Its Genes (Bacillus subtilis NC1 유래 cellulase와 xylanase의 특성 규명 및 효소 유전자의 규명)

  • Park, Chang-Su;Kang, Dae-Ook;Choi, Nack-Shick
    • Journal of Life Science
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    • v.22 no.7
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    • pp.912-919
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    • 2012
  • A Bacillus sp. strain producing celluase and xylanase was isolated from environmental soil with LB agar plate containing carboxymethylcellulose (CM-cellulose) and beechwood xylan stained with trypan blue as substrates, respectively. Based on the 16S rRNA gene sequence and API 50 CHL test, the strain was identified as B. subtilis and named B. subtilis NC1. The cellulase and xylanase from B. subtilis NC1 exhibited the highest activities for CM-cellulose and beechwood xylan as substrate, respectively, and both enzymes showed the maximum activity at pH 5.0 and $50^{\circ}C$. We cloned and sequenced the genes for cellulase and xylanase from genomic DNA of the B. subtilis NC1 by the shot-gun cloning method. The cloned cellulase and xylanase genes consisted of a 1,500 bp open reading frame (ORF) encoding a 499 amino acid protein with a calculated molecular mass of 55,251 Da and a 1,269 bp ORF encoding a 422 amino acid protein with a calculated molecular mass of 47,423 Da, respectively. The deduced amino acid sequences from the genes of cellulase and xylanase showed high identity with glycosyl hydrolases family (GH) 5 and 30, respectively.