• Title/Summary/Keyword: pE194

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Genetic Transformation of Biocontrol Agent Bacillus sp, YBL-7 by Plasmid pE194 (생물방제균 Bacillus sp. YBL-7의 형질전환조건)

  • 한길환;정병곤;김상달
    • Microbiology and Biotechnology Letters
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    • v.20 no.4
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    • pp.384-389
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    • 1992
  • Bacillus sp. YBL-7 which had been isolated from ginseng root-rot suppressive soil was able to antagonize Fusarium solani causing ginseng root-rot by their antibiotic substance. In order to develop multifunctional antagonist on Bacillus sp. YBL-7 as a biocontrol agent against Fusarium salam', optimal conditions for protoplast transformation system of Bacillus sp.YBL-7 by the vector plasmid pE194 were investigated. The protoplasts of Bacillus sp. YBL-7 were obtained at best efficiency by treatment with 200${\mu}g$/ml of lysozyme in the pH 7.0 of SMM buffer for 90 minutes at $40^{\circ}C$. The cell wall of the protoplast was regenerated on the agar plate containing 1.2% agar and 0.7 M mannitol. Under the best condition for protoplast formation and regeneration, the optimal transformation was achieved with 40% polyethylene glycol (M.W. 4000) treatment for 10minutes. The vector plasmid pE194 showed the best transformation frequency at 5$\mu$g/ml of final concentration. The pE194 was very stable over 80% in the transformants.

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Replication origin (ori) of R-plasmid pSBK203 Isolated from Staphylococcus aureus DHI (Staphylococcus aureus DH1에서 분리한 R-plasmid pSBK203상의 복제개시 부위 ori에 관한 연구)

  • Min, Kyung-Il;Byeon, Woo-Hyeon
    • Korean Journal of Microbiology
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    • v.32 no.3
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    • pp.186-191
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    • 1994
  • The origin of the leading strand replication (ori) and of lagging strand replication (M-O) of R-plasmid pSBK203 was identified and its base sequence was determined. About 50 bp of ori sequence residues overlapped with the structural gene of rep. Sequence comparison reveals that pSBK-ori shares obvious identities with those of pT181 family and consists of two regions, one is conserved and the other is variable region. Of two palindrome sequence located one after another in upstream region of rep gene, palA' instead of palA which shares sequence homology with diverse family of plasmids such as pOX6, pC194, and pE194 seems to act as a signal for conversion of primarily replicated ssDNA to dsDNA (minus origin (M-O)).

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Purification and Characterization of Cop, a Protein Involved in the Copy Number Control of Plasmid pE194

  • Kwak, Jin-Hwan;Kim, Jung-Ho;Kim, Mu-Yong;Choi, Eung-Chil
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.291-297
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    • 1998
  • Cop protein has been overexpressed in Escherichia coli using a T7 RNA polymerase system. Purification to apparent homogeneity was achieved by the sequential chromatography on ion exchange, affinity chromatography, and reverse phase high performance liquid chromatography system. The molecular weight of the purified Cop was estimated as 6.1 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). But the molecular mass of the native state Cop was shown to be 19 kDa by an analytical high performance size exclusion chromatography, suggesting a trimer-like structure in 50 mM Tris-HCI buffer (pH 7.5) containing 100 mM NaCl. Cop protein Was calculated to contain $39.1% {\alpha}-helix, 16.8% {\beta}-sheet$, 17.4% turn, and 26.8% random structure. The DNA binding property of Cop protein expressed in E. coli Was preserved during the expression and purification process. The isoelectric point of Cop was determined to be 9.0. The results of amino acid composition analysis and N-terminal amino acid sequencing of Cop showed that it has the same amino acid composition and N-terminal amino acid sequence as those deduced from its DNA sequence analysis, except for the partial removal of N-terminal methionine residue by methionyl-aminopeptidase in E. coli.

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Nucleotide Sequence and Properties of Macrolide-Lincosamide-Streptogramin B Resistance Gene from Staphylococcus aureus DH1 (Staphylococcus aureus DH1에서 분리된 Macrolide-Lincosamide-Streptogramin B 계열 항생물질에 대한 저항성 인자의 특성과 염기서열)

  • 권동현;박승문;윤권상;변우현
    • Korean Journal of Microbiology
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    • v.28 no.1
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    • pp.27-34
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    • 1990
  • Two macrolide-lincosamide-streptogramin B (MLS) antibiotic resistance genes, one expressed inducibly and the other expressed constitutively were recognized from a single Staphylococcus aureus DH1 strain. The inducible MLS resistance gene was isolated and cloned from the R-plasmid pDE1(7.4kb) and the constitutive gene was from chromosomal DNA. Base sequence of the inducible MLS resistance gene (1.2kb) was determined and found as same that of pE194. The restriction map of the cloned constitutive MLS resistance gene was compared with that of the inducible gene. Two genes have same restriction map except leader region. In the constitutive gene there is no leader region which is doing major role in inducible expression.

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Transformation of Bacillus subtilis Protoplast by Recombinant Plasmid DNA (재조합 Plasmid DNA에 의한 Bacillus subtilis의 형질전환)

  • Kim, Sang-Dal;John Spizizen
    • Microbiology and Biotechnology Letters
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    • v.13 no.4
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    • pp.345-348
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    • 1985
  • Recombinant chimeric plasmid constructed with Xba I digested pUBl10 and -pE194 was transformed by polyethylene glycol induced protoplast transformation system into Bacillus subtilis BR 151 on the mannitol regeneration media, and two genes of antibiotics resistance were expressed simultaneously in the transfromant. Transformation frequency of the recombinant plasmid was 6.5 $\times$ 10$^{-5}$ on the mannitol regeneration agar plate containing neomycin and erythromycin. The replication of recombinant plasmid in the recipient cells was confirmed by the alkaline extraction method and agarose gel electrophoresis.

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Indium Tin Oxide Based Reflector for Vertical UV LEDs (자외선 수직형 LED 제작을 위한 Indium Tin Oxide 기반 반사전극)

  • Jung, Ki-Chang;Lee, Inwoo;Jeong, Tak;Baek, Jong Hyeob;Ha, Jun-Seok
    • Korean Journal of Materials Research
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    • v.23 no.3
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    • pp.194-198
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    • 2013
  • In this paper, we studied a p-type reflector based on indium tin oxide (ITO) for vertical-type ultraviolet light-emitting diodes (UV LEDs). We investigated the reflectance properties with different deposition methods. An ITO layer with a thickness of 50 nm was deposited by two different methods, sputtering and e-beam evaporation. From the measurement of the optical reflection, we obtained 70% reflectance at a wavelength of 382 nm by means of sputtering, while only 30% reflectance resulted when using the e-beam evaporation method. Also, the light output power of a $1mm{\times}1mm$ vertical chip created with the sputtering method recorded a twofold increase over a chip created with e-beam evaporation method. From the measurement of the root mean square (RMS), we obtained a RMS value 1.3 nm for the ITO layer using the sputtering method, while this value was 5.6 nm for the ITO layer when using the e-beam evaporation method. These decreases in the reflectance and light output power when using the e-beam evaporation method are thought to stem from the rough surface morphology of the ITO layer, which leads to diffused reflection and the absorption of light. However, the turn-on voltage and operation voltage of the two samples showed identical results of 2.42 V and 3.5 V, respectively. Given these results, we conclude that the two ITO layers created by different deposition methods showed no differences in the electric properties of the ohmic contact and series resistance.

Identification of Polymorphisms in CYP2E1 Gene and Association Analysis among Chronic HBV Patients

  • Chun, Ji-Yong;Park, Byung-Lae;Cheong, Hyun-Sub;Kim, Jason-Y.;Park, Tae-Joon;Lee, Jin-Sol;Lee, Hyo-Suk;Kim, Yoon-Jun;Shin, Hyoung-Doo
    • Genomics & Informatics
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    • v.7 no.4
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    • pp.187-194
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    • 2009
  • Cytochrome P450 2E1 (CYP2E1) is a member of the cytochrome P450 superfamily, and it is a key enzyme responsible for the metabolic activation of many smallmolecular-weight compounds such as alcohol, which is classified as a human carcinogen. In this study, we identified 19 single nucleotide polymorphisms (SNPs) in CYP2E1 in Korean population. In these SNPs, we examined possible genetic association of CYP2E1 polymorphisms with HBV clearance and the risk of hepatocellular carcinoma (HCC). Five common polymorphic sites were selected, CYP2E1 polymorphisms at rs381-3867, rs3813870, rs2070673, rs2515641 and rs2480257, considering their allele frequencies, haplotype-tagging status and LDs for genotyping in larger-scale subjects (n=1,092). Statistical analysis demonstrated that CYP2E1 polymorphisms and haplotypes show no significant association with HBV clearance, HCC occurrence and onset age of HCC (p>0.05). Previous studies, however, have shown contradictory findings on associations of CYP2E1 polymorphisms with CYP2E1 activities and HCC risk. Comparing the contrasting results of previous researches suggest that CYP2E1 polymorphism is associated with CYP2E1 activity induced by ethanol, but is not directly associated with HCC risk. CYP2E1 variation/haploype information identified in this study will provide valuable information for future studies on CYP2E1.

Change in Community Composition and Soil Carbon Stock Along Transitional Boundary in a Sub-Tropical Forest of Garhwal Himalaya

  • Kumar, Munesh;Kumar, Manish;Saleem, Sajid;Prasad, Sunil;Rajwar, G.S.
    • Journal of Forest and Environmental Science
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    • v.29 no.3
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    • pp.194-199
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    • 2013
  • The aim of the present study was to assess the effect of transitional boundary on community composition and soil carbon stock. Five vegetation types were recognized horizontally along the transitional strip based on the dominance of tree species i.e., Pure Anogeissus latifolia forest (P.AL), mixed Pinus roxburghii and Lannea coromandelica forest (M.PR&LC), pure Pinus roxburghii forest (P.PR), mixed Pinus roxburghii and Lannea coromandelica (M.PR&LC) and pure Anogeissus latifolia forest (P.AL). The results revealed that Anogeissus latifolia was reported dominant tree in the outer transitional boundaries of the forest, which reduced dominance of trees towards middle where Pinus roxburghii was found dominant. The soil carbon stock was reported higher in the Anogeissus latifolia dominant forest and reduced with the dominance of Pinus roxburghii in the middle site. Both the species are growing close to one another and competing for survival, but the aggressive nature of Anogeissus latifolia particular in this region may change new growth of Pinus roxburghii and will enhance soil carbon stock. But high anthropogenic pressure on Anogeissus latifolia tree species could be limited chance to further its flourish.

The Effect of Self-leadership and Communication Barriers on Nursing Performance in Hospital Nurses (병원간호사의 셀프리더십, 의사소통 장애가 간호업무성과에 미치는 영향)

  • Cho, Jeong Lim
    • The Journal of the Convergence on Culture Technology
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    • v.5 no.2
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    • pp.239-246
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    • 2019
  • Purpose: The purpose of this study was to identify factors influencing nursing performance in hospital nurses. Methods: The participants were 194 nurses working in one general hospitals in B cities. Data were collected from January 30 to March 30, 2018. SPSS/WIN 18.0 was used for analysis with t-test, ANOVA, $Scheff{\acute{e}}$ test, Pearson correlation coefficients and Stepwise multiple regression. Results: There was a significant correlation between nursing performance and self-leadership (r=.59, p<.001) and communication barriers (r=.35, p<.001). In the multiple regression, self-leadership (${\beta}=.50$, p<.001), position (${\beta}=.15$, p=.011) and communication barriers (${\beta}=.16$, p=.014) were associated with nursing performance. These factors accounted for 37% of the total variance in nursing performance. Conclusion: Considering the results of this study, educational programs to improve self-leadership and communication barriers should be developed and implemented in order to foster nursing performance in hospital nurses.