• Title/Summary/Keyword: High-isolation

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Phylogenetic Relationship and DNA Polymorphism of Boleophthalmus pectinirostris and Scartelaos gigas (Teleostei: Gobiidae) of Korea (한국산 짱뚱어(Boleophthalmus pectinirostris)와 남방짱뚱어(Scartelaos gigas) (Gobiidae)의 분자유전학적 계통연관과 DNA 다형화)

  • Choi, Ki Ho;Chung, Ee Yung;Park, Gab Man
    • Korean Journal of Ichthyology
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    • v.25 no.3
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    • pp.149-156
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    • 2013
  • Phylogenetic relationships and DNA polymorphism among local populations of two Korean gobiidae species: Boleophthalmus pectinirostris and Scartelaos gigas were investigated based on 12S and 16S mitochondrial DNA and mitochondrial cytochrome b DNA sequences. DNA polymorphisms of B. pectinirostris between Suncheon and Gunsan populations were 100% identity from 434 bp segment of 12S rRNA gene and from 444 bp segment of mitochondrial cytochrome b genes, and 99.6% (2 bp different) identity from 484 bp segments of 16S rRNA genes. These results indicated the long period of geographic isolation between two populations of B. pectinirostris in Korea caused such high degrees of DNA polymorphisms. Based on the phylogenetic tree constructed from the two gobiid species in Korea, two genetically distinct groups of B. pectinirostris and S. gigas groups were recognized.

A Study of Ni-resistant bacteria isolated from gingival crevicular fluid on the patients wearing Ni-Cr alloy prosthesis (In terms of molecular biological aspects) (니켈-크롬 합금 보철물 주위 치은열구 내에서 발견된 니켈 내성 균주에 관한 분자생물학적 연구)

  • Chae, Young-Ah;Woo, Yi-Hyung;Choi, Boo-Byung;Choi, Dae-Gyun;Lee, Sung-Bok;Kwon, Kung-Rock
    • The Journal of Korean Academy of Prosthodontics
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    • v.37 no.6
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    • pp.741-755
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    • 1999
  • As a material of metal-ceramic prosthesis, nickel as a form of Ni-Cr alloy has been used for many dental prostheses in many cases. However, several problems in use of the alloy have been revealed (ex : tissue stimulation, skin allergy, hypersensitivity, cytotoxicity and carcinogenecity). Little is known about nickel with respect to the relationship between Ni-prosthesis and gaining of Ni-resistance in oral microorganisms. The present study was undertaken to check wheather use of Ni-prosthesis leads to occurrence of Ni-resistant microorganisms. So this study may suggest the possible relationships between the oral microorganisms and nickel-resistance in oral environment. Bacteria were isolated from the gingival crevicular fluid on the pateints wearing Ni-Cr prosthesis. The isolated bacteria were tested for their Ni-resistance in nickel containing media at different concentration from 3mM to 110mM. E. coli HB101 was used as control. The Ni-resistant bacteria were isolated and biochemically identified. The Ni-resistant bacteria were tested several bio-chemical, molecular-biological tests. Performed tests were ; measuring the growth curve, antibiotic test, growth ability test in liquid media, isolation of the chromosome and plasmid, digestion of DNA by restriction enzyme, electrophoresis of chromosome and plasmid DNA, identification of Ni-resistant genes by the DNA hybridization. The results were as follows: 1) The bacteria isolated from gingival crevicular fluid on the patients wearing Ni-Cr alloy pros-thesis showed nickel-resistance. 2) The isolated microorganisms grew at nickel containing media of high concentrations (60mM-110mM). 3) Based on the biochemical tests, the isolated microorganisms were identified as Enterococcus faecalis(13 cases), Klebsiella pneumoniae(1 case) and Enterobacter gergeviae(1 case). 4) Enterococcus faecalis expressed not only nickel resistance but also the multi-drug resistance to several antibiotics ; chloramphenicol, kanamicin, streptomycin, lincomycin, clindamycin. However, all strain showed the sensitivity against the tetracycline. 5) DNA hybridization result suggest that there is no homology between the previousely known gene of nickel resistance in Klebsiella pneumoniae and chromosomal DNA of Enterococcus faecalis.

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An improvement of real-time polymerase chain reaction system based on probe modification is required for accurate detection of African swine fever virus in clinical samples in Vietnam

  • Tran, Ha Thi Thanh;Dang, Anh Kieu;Ly, Duc Viet;Vu, Hao Thi;Hoang, Tuan Van;Nguyen, Chinh Thi;Chu, Nhu Thi;Nguyen, Vinh The;Nguyen, Huyen Thi;Truong, Anh Duc;Pham, Ngoc Thi;Dang, Hoang Vu
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.10
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    • pp.1683-1690
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    • 2020
  • Objective: The rapid and reliable detection of the African swine fever virus (ASFV) plays an important role in emergency control and preventive measures of ASF. Some methods have been recommended by FAO/OIE to detect ASFV in clinical samples, including realtime polymerase chain reaction (PCR). However, mismatches in primer and probe binding regions may cause a false-negative result. Here, a slight modification in probe sequence has been conducted to improve the qualification of real-time PCR based on World Organization for Animal Health (OIE) protocol for accurate detection of ASFV in field samples in Vietnam. Methods: Seven positive confirmed samples (four samples have no mismatch, and three samples contained one mutation in probe binding sites) were used to establish novel real-time PCR with slightly modified probe (Y = C or T) in comparison with original probe recommended by OIE. Results: Both real-time PCRs using the OIE-recommended probe and novel modified probe can detect ASFV in clinical samples without mismatch in probe binding site. A high correlation of cycle quantification (Cq) values was observed in which Cq values obtained from both probes arranged from 22 to 25, suggesting that modified probe sequence does not impede the qualification of real-time PCR to detect ASFV in clinical samples. However, the samples with one mutation in probe binding sites were ASFV negative with OIE recommended probe but positive with our modified probe (Cq value ranked between 33.12-35.78). Conclusion: We demonstrated for the first time that a mismatch in probe binding regions caused a false negative result by OIE recommended real-time PCR, and a slightly modified probe is required to enhance the sensitivity and obtain an ASF accurate diagnosis in field samples in Vietnam.

Anti-metastatic Effects on B16F10 Melanoma Cells of Extracts and Two Prenylated Xanthones Isolated from Maclura amboinensis Bl. Roots

  • Siripong, Pongpun;Rassamee, Kitiya;Piyaviriyakul, Suratsawadee;Yahuafai, Jantana;Kanokmedhakul, Kwanjai
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3519-3528
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    • 2012
  • Inhibitory effects of Maclura amboinenesis Bl, one plant used traditionally for the treatment of cancers, on metastatic potential of highly metastatic B16F10 melanoma cells were investigated in vitro. Cell proliferation was assessed using the MTT colorimetric assay. Details of metastatic capabilities including invasion, migration and adhesion of B16F10 melanoma cells were examined by Boyden Chamber invasion and migration, scratch motility and cell attachment assays, respectively. The results demonstrated that n-hexane and chloroform extracts exhibited potent anti-proliferative effects (p<0.01), whereas the methanol and aqueous extracts had less pronounced effects after 24 h exposure. Bioactivity-guided chromatographic fractionation of both active n-hexane and chloroform extracts led to the isolation of two main prenylated xanthones and characterization as macluraxanthone and gerontoxanthone-I, respectively, their structures being identified by comparison with the spectral data. Interestingly, both exhibited potent effective effects. At non-toxic effective doses, n-hexane and chloroform extracts (10 and $30{\mu}g/ml$) as well as macluraxanthone and gerontoxanthone-I (3 and $10{\mu}M$) significantly inhibited B16F10 cell invasion, to a greater extent than $10{\mu}m$ doxorubicin, while reducing migration of cancer cells without cellular cytotoxicity. Moreover, exposure of B16F10 melanoma cells to high concentrations of chloroform ($30{\mu}g/ml$) and geratoxanthone-I ($20{\mu}M$) for 24 h resulted in delayed adhesion and retarded colonization. As insights into mechanisms of action, typical morphological changes of apoptotic cells e.g. membrane blebbing, chromatin condensation, nuclear fragmentation, apoptotic bodies and loss of adhesion as well as cell cycle arrest in the G1 phase with increase of sub-G1 cell proportions, detected by Hoechst 33342 staining and flow cytometry were observed, suggesting DNA damage and subsequent apoptotic cell death. Taken together, our findings indicate for the first time that active n-hexane and chloroform extracts as well as macluraxanthone and gerontoxanthone-I isolated from Maclura amboinensis Bl. roots affect multistep of cancer metastasis processes including proliferation, adhesion, invasion and migration, possibly through induction of apoptosis of highly metastatic B16F10 melanoma cells. Based on these data, M. amboinensis Bl. represents a potential candidate novel chemopreventive and/or chemotherapeutic agent. Additionally, they also support its ethno-medicinal usage for cancer prevention and/or chemotherapy.

Isolation and Molecular Characterization of a New CRT Binding Factor Gene from Capsella bursa-pastoris

  • Wang, Xinglong;Liu, Li;Liu, Sixiu;Sun, Xiaoqing;Deng, Zhongxiang;Pi, Yan;Sun, Xiaofen;Tang, Kexuan
    • BMB Reports
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    • v.37 no.5
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    • pp.538-545
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    • 2004
  • A new CRT binding factor (CBF) gene designated Cbcbf25 was cloned from Capsella bursa-pastoris, a wild grass, by the rapid amplification of cDNA ends (RACE). The full-length cDNA of Cbcbf25 was 898 bp with a 669 bp open reading frame (ORF) encoding a putative DRE/CRT (LTRE)-binding protein of 223 amino acids. The predicted CbCBF25 protein contained a potential nuclear localization signal (NLS) in its N-terminal region followed by an AP2 DNA-binding motif and a possible acidic activation domain in the C-terminal region. Bioinformatic analysis revealed that Cbcbf25 has a high level of similarity with other CBF genes like cbf1, cbf2, and cbf3 from Arabidopsis thaliana, and Bncbf5, Bncbf7, Bncbf16, and Bncbf17 from Brassica napus. A cold acclimation assay showed that Cbcbf25 was expressed immediately after cold triggering, but this expression was transient, suggesting that it concerns cold acclimation. Our study implies that Cbcbf25 is an analogue of other CBF genes and may participate in cold-response, by for example, controlling the expression of cold-regulated genes or increasing the freezing tolerance of plants.

Human Being in the Contemporary Society (도시적 인간상 연구 - 본인 작품을 중심으로-)

  • 박성원
    • Proceedings of the Korea Contents Association Conference
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    • 2004.05a
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    • pp.553-561
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    • 2004
  • It works as intermediation of communication of publics these days. Since 20 century, We, Koreans, have established new chaotic multi culture with traditional Korean culture and other different culture from everywhere. Meanwhile, we occupied the most powerful semi-conduct and IT indusry. Within those circumstance, people feel very confused in political, cultural and social aspect. The society armed with economy and popularization promotes material satisfaction with this potential possibility of anonymous masses. However, it results to cause loneliness, isolation, alienation, anonymity, non individuality and commodity of culture. In my work, such phenomenon reveals through human character in a city. People are exposed culture of consumption and surrounded and tempted by all those artificial and superficial atmosphere. Human are possessed and exposed to attractive products and visual images. Finally they make themselves stuck in their case of this world. People lose their own identify and shape of bodies. That is our portrait, who are living this moment. Also, this is a symbol that destroys this modern society. As a result, 1 consider such aspects through those elements above to think how to keep and rethink our identity and what to do for this world.

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Isolation and Characterization of Dextrans Produced by Leuconostoc sp. strain JYY4 from Fermented Kimchi

  • Gu, Ji-Joong;Ha, Yoo-Jin;Yoo, Sun-Kyun
    • Journal of the Korean Applied Science and Technology
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    • v.32 no.4
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    • pp.758-766
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    • 2015
  • Dextran is a generic term for a bacterial exopolysaccharide synthesized from sucrose and composed of chains of D-glucose units connected by ${\alpha}$-1,6-linkages by using dextransucrases. Dextran could be used as vicosifying, stabilizing, emulsifying, gelling, bulking, dietary fiber, prebiotics, and water holding agents. We isolated new strain capable of producing dextran from Korean traditional kimchi and identified as Leuconostoc sp. strain JYY4. Batch fermentation was conducted in bioreactor with a working volume of 3 L. The media was MMY and 15% (w/v) sucrose. Mineral medium consisted of $3.0g\;KH_2PO_4$, $0.01g\;FeSO_4$, $H_2O$, $0.01g\;MnSO_4$, $4H_2O$, $0.2g\;MgSO_4\;7H_2O$, 0.01 g NaCl, $0.05g\;CaCl_2$ per 1 liter deionized water. The pH of media was initially adjusted to 6.0. The inoculation rate was 1.0% (v/v) of the working volume. Temperature was maintained at $28^{\circ}C$. The agitation rate was 100 rpm. The production pattern of dextran was associated with the cell growth. After 24 hr dextran reached its highest concentration of 59.4 g/L. The sucrose was consumed completely after 40 hr. Growth reached stationery phase when sucrose became limiting, regardless of the presence of fructose or mannitol. When the specific growth rate was 0.54 hr-1, utilization averaged 5.8 g/L-hr. The yield and productivity of dextran were 80% and 2.0 g/L-hr, respectively. Dextrans produced by were separated to two different size by an alcohol fraction method. The size of high molecular weight dextran (45% alcohol, v/v), less soluble dextran, was between MW 500,000 and 2,000,000. Soluble dextran (55% alcohol, v/v) was between 70,000 and 150,000. The molecular weight average of total dextran (70% alcohol, v/v) was between 150,000 to 500,000. The enzymatic hydrolyzates of total dextran of ATCC 13146 showed branched dextrans by Penicillium dextranase contained of glucose, isomaltose, isomaltotriose, and isomaltooligosaccharides greater than DP4 (degree of polymerization) that had branch points. Compounds greater than DP4 were branched isomaltooligosaacharides. Hydrolysates by the Lipomyces dextranase produced the same composition of oligosaccharides as those by Penicillin dextranase.

Citric acid Fermentation by Mutant Strain of Candida lipolytica (Candida lipolytica 변이주에 의한 구연산발효)

  • 전효곤;성낙기;박석규
    • Microbiology and Biotechnology Letters
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    • v.13 no.3
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    • pp.245-250
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    • 1985
  • In order to increase citric acid productivity. several attempts were made; isolation and characterization of the mutant strain produced citric acid in a high yield, citric acid fermentation in a medium containing relatively higher amount of glucose and citric acid production by the use of semicontinuous ceil recycle system. By the treatment of Candide lipolytica S-109 with NTG, a mutant J-24 was selected as the highest producer of citric acid among the strains formed larger CaCO$_3$ lytic zone. it produced 72g/1 citric acid in 10% glucose medium. Because mutant J-24 produced 85g/l citric acid and showed 53% yield in 16% glucose medium, several factors were adjusted to increase the yield in 16% glucose medium. 0.8-1.0$\times$10$^{-3}$ P/C ratio, 0.15% urea, 0.25% yeast extract were suitable at citric acid production in 16% glucose medium. Under this condition, J-24 strain produced 93g/l citric acid and showed 58% yield. Semicontinuous cell recycle system was used to protons the effective production phase, to minimize the product inhibition and to shorten the lag phase. The productivity of semicontinuous cell recycle system was 0.79g/l h while that of batch system was 0.53g/l.h

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Lysine Production by Thialysine Resistant Mutant of Candida utilis ( I ) - Isolation of High Lysine Excreting Mutant of Candida utilis - (Candida utilis의 Thialysine 내성맥리주에 의한 Lysine생산 ( I ) -Candida utilis의 Lysine을 생산하는 Thialysine 내성맥리주의 분리-)

  • Bang, Byung-Ho;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.11 no.3
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    • pp.175-180
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    • 1983
  • Thialysine significantly inhibited the growth of wild type strain Gondida utilis NCYC-359. In the absence of thialysine, the culture reached stationary phase after 24hr, however, in the presence of 0.5% thialysine, the culture reached stationary phase after 40hr, respectively. Effect of amino acid or vitamin was investigated on recovery of the growth of wild type strain from thialysine inhibition. Glycine, methionine, arginine and tryptophan recovered growth inhibition by thialyslne to some extent. However, vitamins were inert. Especially, lysine at one eighth concentration of thialysine recovered almost fully the growth inhibition. Thialysine resistant mutants were induced from the parent strain of Condida utilis NCYC-359 by NTG treatment. Colonies of thialysine resistant mutants were obtained on agar minimal medium supplemented with 0.1-0.5% thialysine. The frequency of thialysine resistant mutants induced by the first mutation was the highest at 0.1% The wild strain produced no appreciable lysine extracellularly. However, almost thialysine resistant mutants excreted appreciably. Lysine excretion increased after repeated mutation. Finally, of the thialysine resistant mutants induced by NTG, Condida utilis TRN-4006 was obtained. This strain excreted lysine (400$\mu\textrm{g}$/$m\ell$) into the medium with a concomitant decrease of lysine in the intracellular pool.

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Genetic Variation of Two Isolated Relict Populations of Vaccinium uliginosum L. in Korea (들쭉나무 격리잔존 2개 집단의 유전변이)

  • Han, Sang-Don;Hong, Yong-Pyo;Kwon, Hae-Yun;Yang, Byeung-Hoon;Kim, Chan-Soo
    • Journal of Korean Society of Forest Science
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    • v.94 no.4 s.161
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    • pp.209-213
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    • 2005
  • In order to provide the molecular genetic information necessary for conservation of bog whortleberry (Vaccinium uliginosum L), one of the rare species in Korea, I-SSR analysis was performed on two populations on Mt. Halla and Mt. Seorak. A total of 68 I-SSR products were observed, and higher level of genetic diversity was observed in Mt. Halla population (S.I.=0.539) than in the Mt. Seorak population (S.I,=0.401). Level of genetic diversity in this species was relatively higher than those in other rare species analysed with I-SSR marker. From the results of AMOVA, exceptionally large proportion of genetic diversity (33.5%) was resulted from genetic difference between two populations, and only 66.5% of the genetic variation was allocated in common among individuals within each population, compared with the results in other long-lived woody species. This remarkably high degree of genetic heterogeneity existed between Mt. Halla and Mt. Seorak populations might suggest that they might be originated from the independent progenitors before the post glacier ages, respectively, and/or that they undergone random genetic drift respectively due to geographical isolation resulted from dramatic changes in environmental conditions after the post glacier ages.