• Title/Summary/Keyword: Positive Clone

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Phylogenetic Diversity of Bacteria in an Earth-Cave in Guizhou Province, Southwest of China

  • Zhou, Jun-Pei;Gu, Ying-Qi;Zou, Chang-Song;Mo, Ming-He
    • Journal of Microbiology
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    • v.45 no.2
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    • pp.105-112
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    • 2007
  • The objective of this study was to analyze the phylogenetic composition of bacterial community in the soil of an earth-cave (Niu Cave) using a culture-independent molecular approach. 16S rRNA genes were amplified directly from soil DNA with universally conserved and Bacteria-specific rRNA gene primers and cloned. The clone library was screened by restriction fragment length polymorphism (RFLP), and representative rRNA gene sequences were determined. A total of 115 bacterial sequence types were found in 190 analyzed clones. Phylogenetic sequence analyses revealed novel 16S rRNA gene sequence types and a high diversity of putative bacterial community. Members of these bacteria included Proteobacteria (42.6%), Acidobacteria (18.6%), Planctomycetes (9.0 %), Chloroflexi (Green nonsulfur bacteria, 7.5%), Bacteroidetes (2.1%), Gemmatimonadetes (2.7%), Nitrospirae (8.0%), Actinobacteria (High G+C Gram-positive bacteria, 6.4%) and candidate divisions (including the OP3, GN08, and SBR1093, 3.2%). Thirty-five clones were affiliated with bacteria that were related to nitrogen, sulfur, iron or manganese cycles. The comparison of the present data with the data obtained previously from caves based on 16S rRNA gene analysis revealed similarities in the bacterial community components, especially in the high abundance of Proteobacteria and Acidobacteria. Furthermore, this study provided the novel evidence for presence of Gemmatimonadetes, Nitrosomonadales, Oceanospirillales, and Rubrobacterales in a karstic hypogean environment.

Cloning and Functional Expression in Escherichia coli of the Polyhydroxyalkanoate Synthase (phaC) Gene from Alcaligenes sp. SH-69

  • Lee, Il;Nam, Sun-Woo;Rhee, Young-Ha;Kim, Jeong-Yoon
    • Journal of Microbiology and Biotechnology
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    • v.6 no.5
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    • pp.309-314
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    • 1996
  • Alcaligenes sp. SH-69 can synthesize poly(3-hydroxybutyrate-co-3-hydroxyvalerate) from a single carbon source such as glucose. To clone the phaC gene from Alcaligenes sp. SH-69, a polymerase chain reaction was performed using the oligomers synthesized based on the conserved regions of the phaC genes from other bacteria. A PCR product (550 bp) was partially sequenced and the deduced amino acid sequence was found to be homologous to that of the phaC gene from Alcaligenes eutrophus. Using the PCR fragment Southern blotting of Alcaligenes sp. SH-69 genomic DNA digested with several restriction enzymes was carried out. To prepare a partial genomic library, about 5-Kb genomic DNA fragments digested with EcoRI, which showed a positive signal in the Southern blotting, were eluted from an agarose gel, ligated with pUC19 cleaved with EcoRI, and transformed into Escherichia coli. The partial library was screened using the PCR fragment as a probe and a plasmid, named pPHA11, showing a strong hybridization signal was selected. Restriction mapping of the insert DNA in pPHA11 was performed. Cotransformation into E. coli of the plasmid pPHA11 and the plasmid pPHA21 which has phaA and phaB from A. eutrophus resulted in turbid E. coli colonies which are indicative of PHA accumulation. This result tells us that the Alcaligenes sp. SH-69 phaC gene in the pPHA11 is functionally active in E. coli and can synthesize PHA in the presence of the A. eutrophus phaA and phaB genes.

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Comparison of the Virulence-Associated Phenotypes of Five Species of Acinetobacter baumannii Complex

  • Na, In Young;Chung, Eun Seon;Jung, Chang-Yun;Kim, Dae Hun;Shin, Juyoun;Kang, KyeongJin;Kim, Seong-Tae;Ko, Kwan Soo
    • Journal of Microbiology and Biotechnology
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    • v.26 no.1
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    • pp.171-179
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    • 2016
  • In this study, we compared the virulence-associated factors of Acinetobacter baumannii complex species. Sixty-three isolates of five A. baumannii complex species, including 19 A. baumannii, 15 A. nosocomialis, 13 A. seifertii, 13 A. pittii, and 3 A. calcoaceticus isolates, were included in this study. For all isolates, biofilm formation, A549 cell adherence, resistance to normal human serum, and motility were evaluated. A. baumannii complex isolates showed diversity in biofilm formation, A549 cell adherence, and serum resistance, and no strong positive relationships among these virulence characteristics. However, A. seifertii showed relatively consistent virulence-associated phenotypes. In addition, A. baumannii clone ST110 exhibited consistently high virulence-associated phenotypes. Motility was observed in seven isolates, and all four A. baumannii ST110 isolates showed twitching motility. Although some inconsistencies in virulence-associated phenotypes were seen, high virulence characteristics were observed in A. seifertii, which has been mainly reported in Korea and shows high rates of colistin resistance.

Genetic Variability, Correlation and Path Analysis for Yield Components in Korea Domestic Ginger (한국재래생강(韓國在來生薑)에 있어서 수량구성요소(收量構成要素)의 유전변이(遺傳變異), 상관(相關) 및 경로분석(經路分析))

  • Jang, Won Suk;Kim, Jung Sun;Choi, Jae Eul
    • Korean Journal of Agricultural Science
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    • v.24 no.1
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    • pp.6-10
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    • 1997
  • Ninety-four collections of domestic ginger were evaluated for plant hight, leaf number, stem number, leaf length and width, rhizome yield per plant revealed good variability in stem number and rhizome yield per plant. Moderate variation was observed in plant hight, leaf number, leaf length, leaf width and stem diameter. Rhizome yield per plant was positively correlated with six characters, and leaf number, plant height and stem diameter were positive significant correlation with rhizome yield per plant. Path coefficient analysis indicated that stem number, followed by leaf number, stem diameter, had maximum direct effects on rhizome yield per plant. Maximum indirect effect was observed in case of plant height through leaf number. The indirect effects of leaf length and leaf width through leaf number, and plant height, leaf length and leaf width through stem diameter were also observed. From the selection point of view, the characters like plant height, stem number and leaf number per clump may be considered suitable in choosing a good genotype.

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Distribution of Pathogenic Vibrios in the Aquatic Environment Adjacent to Coastal Areas of South Korea and Analysis of the Environmental Factors Affecting Their Occurrence (2016년도 국내 해양환경내 병원성 비브리오균의 분포 및 해양환경인자간의 상관성 분석)

  • Jeong, Young-Il;Myung, Go-Eun;Choi, Eun-Jin;Soh, Sang-Moon;Park, Gi-Jun;Son, Tae-Jong
    • Journal of Environmental Health Sciences
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    • v.44 no.2
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    • pp.133-142
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    • 2018
  • Objectives: The pathogenic Vibrios genus denotes halophilic bacteria that are distributed in aquatic environments, including both sea and freshwater. Vibrio cholerae, Vibrio vulnificus, and Vibrio parahaemolyticus are the most important species since they can be potent human pathogens and leading causes of septicemia, wound infections, and seafood borne gastroenteritis. The recent emergence of a potential pandemic clone, V. cholera serotype O1 and the cholera outbreak in South Korea in 2016 indicates the importance of consistent surveillance of pathogenic Vibrio genus within coastal areas. Methods: The present study was undertaken to determine where and how vibrios live in the aquatic environment adjacent to coastal areas of South Korea. For this survey, a total of 838 samples were obtained at 35 different sites in South Korean coastal areas during the period from January 2016 to December 2016. Pathogenic vibrios was determined using the real-time PCR method, and its clones were isolated using three selective plating media. We also monitored changes in seawater and atmospheric temperature, salinity, turbidity, and hydrogen ion concentration at the collection points. Results: The total isolation rates of V. vulnificus, V. cholera (non-pathogenic, non-O1, non-O139 serogroups), and V. parahaemolyticus from seawater specimens in 2016 were 14.2, 13.48, and 67.06%, respectively. Conclusions: The isolation rates of pathogenic vibrios genus showed a positive correlation with temperature of seawater and atmosphere but were negatively correlated with salinity and turbidity.

Isolation and Nucleotide Sequence Characterization of Novel Cytochrome P450 Hydroxylase Genes from Rare Actinomycetes, Sebekia benihana (희소 방선균 Sebekia benihana 유래 신규 사이토크롬 P450 하이드록실레이즈 유전자군 분리 및 염기서열 특성규명)

  • 박남실;박현주;한규범;김상년;김응수
    • KSBB Journal
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    • v.19 no.4
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    • pp.308-314
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    • 2004
  • A degenerate set of PCR primers based on two conserved regions (heme binding region and oxygen ligand pocket) were designed and successfully applied to amplify DNA fragments of cytochrome P450 hydroxylase (CYP) genes from a rare actinomycetes, S. benihana. The PCR amplified products were employed as a DNA probe to clone the entire CYP genes from S. benihana genomic library. Five different CYP-positive cosmids were isolated by colony hybridization as well as PCR confirmation. The complete nucleotide sequencing of five different CYP genes revealed that each unique CYP showed a significant amino acid homology to previously-known CYP genes involved in streptomycetes secondary metabolism. In addition, four CYP genes (CYP502, CYP503, CYP504, CYP506) were found to be linked to ferredoxin genes in the chromosome, and the CYP503 gene showed the high degree of amino acid similarity to the previously well-characterized CYP105 family in streptomycetes.

Effect of the Mixture of Pueraria lobata and Sorbus commixta Extract on the Alcohol-induced Hangover in Rats

  • Hong, Se Chul;Yoo, Ji Hyun;Oh, Myeong Hwan;Lee, Hwan;Park, Young Sik;Parthasarathi, Shanmugam;Park, Jong Dae;Pyo, Mi Kyung
    • Natural Product Sciences
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    • v.21 no.2
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    • pp.98-103
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    • 2015
  • Pueraiae Radix (PR), Pueratia Folium (PF) and Sorbus commixta (SC) mixture, namely GS-SP (PR (1)/PF (2)/SC (0.5): v/v/v) was developed as hangover-relieving elixir and its effects on alcoholic metabolism have been investigated. The enzymatic activity of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) of GS-SP was shown higher than those of single treatment with PR, PL, SC, and the positive control group (YM-808). The survival rate of mouse liver cell line NCTC clone 1469 in the presence of acetaldehyde was 30.6, 22.2, and 8.7% at the GS-SP dosage level of 50, 100, and 200 μg/mL respectively. Different concentrations of 50, 100 and 200 mg/kg of GS-SP showed efficient activity for ADH and ALDH than YM-808 in rat fed with 25% ethanol. The levels of blood alcohol and acetaldehyde after oral administration of 200 mg/kg of GS-SP showed efficient activity of 11.7% and 37% than those of YM-808. These results have been supported to the potential for GS-SP to serve as an excellent potential in providing hangover relief and liver protection.

Screening and Characterization of a Novel Cellulase Gene from the Gut Microflora of Hermetia illucens Using Metagenomic Library

  • Lee, Chang-Muk;Lee, Young-Seok;Seo, So-Hyeon;Yoon, Sang-Hong;Kim, Soo-Jin;Hahn, Bum-Soo;Sim, Joon-Soo;Koo, Bon-Sung
    • Journal of Microbiology and Biotechnology
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    • v.24 no.9
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    • pp.1196-1206
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    • 2014
  • A metagenomic fosmid library was constructed using genomic DNA isolated from the gut microflora of Hermetia illucens, a black soldier fly. A cellulase-positive clone, with the CS10 gene, was identified by extensive Congo-red overlay screenings for cellulase activity from the fosmid library of 92,000 clones. The CS10 gene was composed of a 996 bp DNA sequence encoding the mature protein of 331 amino acids. The deduced amino acids of CS10 showed 72% sequence identity with the glycosyl hydrolase family 5 gene of Dysgonomonas mossii, displaying no significant sequence homology to already known cellulases. The purified CS10 protein presented a single band of cellulase activity with a molecular mass of approximately 40 kDa on the SDS-PAGE gel and zymogram. The purified CS10 protein exhibited optimal activity at $50^{\circ}C$ and pH 7.0, and the thermostability and pH stability of CS10 were preserved at the ranges of $20{\sim}50^{\circ}C$ and pH 4.0~10.0. CS10 exhibited little loss of cellulase activity against various chemical reagents such as 10% polar organic solvents, 1% non-ionic detergents, and 0.5 M denaturing agents. Moreover, the substrate specificity and the product patterns by thin-layer chromatography suggested that CS10 is an endo-${\beta}$-1,4-glucanase. From these biochemical properties of CS10, it is expected that the enzyme has the potential for application in industrial processes.

Plagiarism Detection Using Dependency Graph Analysis Specialized for JavaScript (자바스크립트에 특화된 프로그램 종속성 그래프를 이용한 표절 탐지)

  • Kim, Shin-Hyong;Han, Tai-Sook
    • Journal of KIISE:Software and Applications
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    • v.37 no.5
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    • pp.394-402
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    • 2010
  • JavaScript is one of the most popular languages to develope web sites and web applications. Since applicationss written in JavaScript are sent to clients as the original source code, they are easily exposed to plagiarists. Therefore, a method to detect plagiarized JavaScript programs is necessary. The conventional program dependency graph(PDG) based approaches are not suitable to analyze JavaScript programs because they do not reflect dynamic features of JavaScript. They also generate false positives in some cases and show inefficiency with large scale search space. We devise a JavaScript specific PDG(JS PDG) that captures dynamic features of JavaScript and propose a JavaScript plagiarism detection method for precise and fast detection. We evaluate the proposed plagiarism detection method with experiment. Our experiments show that our approach can detect false-positives generated by conventional PDG and can prune the plagiarism search space.

A case of transitional carcinoma in the nasal cavity of a dog (개의 비강 내 이행암종 증례)

  • Kang, Hwa-Jung;Kang, Sang-Chul;Jung, Ji-Youl;Jo, Suk-Hee;Roh, In-Soon;Lee, Joo-Myung;Cheong, Jongtae;Kim, Jung-Hun;An, Min-Chan;Bae, Jong-Hee;Kim, Jae-Hoon
    • Korean Journal of Veterinary Research
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    • v.48 no.3
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    • pp.305-310
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    • 2008
  • A 10-year-old female Yorkshire terrier with the clinical signs of nasal swelling, epistaxis and nasal discharge was presented to the Veterinary Teaching Hospital in the Cheju National University. Abnormal nasal mass was detected in physical examination and radiographic findings. After surgical excision, the sample of nasal mass was referred to Pathology Department of Veterinary Medicine. Grossly, the mass was soft, friable, and $2.5{\times}4cm$ cm in size. Histopathologically, the mass was composed of mediumsized non-keratinizing columnar to polyhedral cells arranged in anastomosing ribbon and large nest. It has complex in-folding of thick epithelial layers separated by fibrovascular septa. Tumor cells showed characteristic palisading arrangement of columnar cells, and perpendicularly distributed to the basement membrane. The cells had pale basophilic cytoplasm, oval nucleus and one or more nucleoli, and indistinct cellular border. Many tumor cell emboli were presented in lymphatics. Immunohistochemistry revealed that tumor cells were cytokeratin (CK) 19 and CK clone MNF116 positive and but CK7 and CK high molecular weight negative. Based on the gross, histopathologic, and immunohistochemical findings, the mass was diagnosed as transitional carcinoma in nasal cavity. In our best knowledge, this is the first report of transitional carcinoma originated from transitional zone of canine nasal cavity in Korea.