• Title/Summary/Keyword: envZ

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CheY-OmpR Hybrid Protein Acting on the Osmoregulatory System (CheY-OmpR 혼성 단백질의 삼투조절효과)

  • 고민수;박찬규
    • Korean Journal of Microbiology
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    • v.33 no.2
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    • pp.118-124
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    • 1997
  • In the previous study(6), we constructed the CheY-OmpR hybrid, Chp, which affects the expressions of ompF and om pC genes. Here we further characterize these effects and present the regulatory mechanism based on in vivo and in vitro data. Although Chp retained the sequence-specific DNA-binding ability, it was not possible to enhance transcriptional activity, suggesting that it may act as a competitive inhibitor to OmpR. The DNA-binding affinity of Chp was not modulated by phosphorylation of its Che Y portion. Chp was able to increase ompR transcription. FurthemlOre, it was found that the wild-type OmpR also exerts the same effect, which is also eOlltrolled by changes in medium osmolarity and in EnvZ activity.

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Construction of Methanol-Sensing Escherichia coli by the Introduction of a Paracoccus denitrificans MxaY-Based Chimeric Two-Component System

  • Ganesh, Irisappan;Vidhya, Selvamani;Eom, Gyeong Tae;Hong, Soon Ho
    • Journal of Microbiology and Biotechnology
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    • v.27 no.6
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    • pp.1106-1111
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    • 2017
  • Escherichia coli was engineered to sense methanol by employing a chimeric two-component system (TCS) strategy. A chimeric MxaY/EnvZ (MxaYZ) TCS was constructed by fusing the Paracoccus denitrificans MxaY with the E. coli EnvZ. Real-time quantitative PCR analysis and GFP-based fluorescence analysis showed maximum transcription of ompC and the fluorescence at 0.01% of methanol, respectively. These results suggested that E. coli was successfully engineered to sense methanol by the introduction of chimeric MxaYZ. By using this strategy, various chimeric TCS-based bacterial biosensors can be constructed and used for the development of biochemical-producing recombinant microorganisms.

Comparison of Methods Predicting VS30 from Shallow VS Profiles and Suggestion of Optimized Coefficients (얕은 심도 VS주상도를 활용한 VS30 예측 방법론 비교 및 최적 계수 제시)

  • Choi, Inhyeok;Kwak, Dongyoup
    • Journal of the Korean Geotechnical Society
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    • v.36 no.3
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    • pp.15-23
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    • 2020
  • Ground motion models predicting intensity measures on surface use a time-averaged shear wave velocity, VS30, as a key variable simulating site effect. The VS30 can be directly estimated from VS profiles if the profile depth (z) is greater than or equal to 30 m. However, some sites have VS profiles with z < 30 m. In this case VS30 can be predicted using extension models. This study proposes new coefficient sets for existing prediction equations using 297 Korea VS profiles. We have collected VS profiles from KMA and Geoinfo database. Fitting six existing methods to data, we suggest new coefficients for each method and evaluate their performance. It turns out that if z ≥ 15 m, the standard deviation (σ) of residual in log10 is 0.061, which indicates that the estimated VS30 is nearly accurate. If z < 15 m, the σ keeps increasing up to 0.1 for z = 5 m, so we caution the use of models at very low z. Nonetheless, we recommend investigating up to 30 m depth for VS30 calculation if possible.

Z-DNA-Containing Long Terminal Repeats of Human Endogenous Retrovirus Families Provide Alternative Promoters for Human Functional Genes

  • Lee, Du Hyeong;Bae, Woo Hyeon;Ha, Hongseok;Park, Eun Gyung;Lee, Yun Ju;Kim, Woo Ryung;Kim, Heui-Soo
    • Molecules and Cells
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    • v.45 no.8
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    • pp.522-530
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    • 2022
  • Transposable elements (TEs) account for approximately 45% of the human genome. TEs have proliferated randomly and integrated into functional genes during hominoid radiation. They appear as right-handed B-DNA double helices and slightly elongated left-handed Z-DNAs. Human endogenous retrovirus (HERV) families are widely distributed in human chromosomes at a ratio of 8%. They contain a 5'-long terminal repeat (LTR)-gag-pol-env-3'-LTR structure. LTRs contain the U3 enhancer and promoter region, transcribed R region, and U5 region. LTRs can influence host gene expression by acting as regulatory elements. In this review, we describe the alternative promoters derived from LTR elements that overlap Z-DNA by comparing Z-hunt and DeepZ data for human functional genes. We also present evidence showing the regulatory activity of LTR elements containing Z-DNA in GSDML. Taken together, the regulatory activity of LTR elements with Z-DNA allows us to understand gene function in relation to various human diseases.

Morphological Features of Coleosporium xanthoxyli and Its Alternate Host in Korea (산초나무 잎녹병균의 중간기주 및 형태학적 특징)

  • Lee, S.K.;Lee, K.H.;Lee, C.K.;Kim, D.Y.;Hwang, J.H.
    • Research in Plant Disease
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    • v.10 no.4
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    • pp.279-284
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    • 2004
  • A rust fungus has caused a serious early defoliation of Zanthoxylum schinifolium during growing seasons every year at the plantations located at Hadong and Jinju, Kyeongsangnam-Do in Korea. In order to identify the rust fungus and clarify its life cycle in Korea, aeciospores from Pinus thunbergii were artificially inoculated on the leaves of Z. schinifolium. Uredinial stage was successively formed on the leaves of Z. schinifolium. Based on the artificial inoculation test and on the morphological features of the dried specimens collected from P. thunbergii and Z. schinifolium, this rust fungus was identified as Coleosporium xanthoxyli. Morphological features of aecial and uredinial stages of the species were described. The first symptom of the infection was developed from later June to early July. And leaf infection ratio was 17.8%-58.7% during August at Hadong and Jinju regions of Kyeongsangnam-Do in Korea.

Investigation of Stiffness Characteristics of Subgrade Soils under Tracks Based on Stress and Strain Levels (응력 및 변형률 수준을 고려한 궤도 흙노반의 변형계수 특성 분석)

  • Lim, Yujin;Kim, DaeSung;Cho, Hojin;Sagong, Myoung
    • Journal of the Korean Society for Railway
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    • v.16 no.5
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    • pp.386-393
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    • 2013
  • In this study, the so-called repeated plate load bearing test (RPBT) used to get $E_{v2}$ values in order to check the degree of compaction of subgrade, and to get design parameters for determining the thickness of the trackbed foundation, is investigated. The test procedure of the RPBT method is scrutinized in detail. $E_{v2}$ values obtained from the field were verified in order to check the reliability of the test data. The $E_{v2}$ values obtained from high-speed rail construction sites were compared to converted modulus values obtained from resonant column (RC) test results. For these tests, medium-size samples composed of the same soils from the field were used after analyzing stress and strain levels existing in the soil below the repeated loading plates. Finite element analyses, using the PLAXIS and ABAQUS programs, were performed in order to investigate the impact of the strain influence coefficient. This was done by getting newly computed $I_z$ to get the precise strain level predicted on the subgrade surface in the full track structure; under wheel loading. It was verified that it is necessary to use precise loading steps to construct nonlinear load-settlement curves from RPBT in order to get correct $E_{v2}$ values at the proper strain levels.

Engineering of Recombinant Escherichia coli Towards Methanol Sensing Using Methylobacterium extroquens Two-component Systems

  • Selvamani, Vidhya;Ganesh, Irisappan;Chae, Sowon;Maruthamuthu, Murali kannan;Hong, Soon Ho
    • Microbiology and Biotechnology Letters
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    • v.48 no.1
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    • pp.24-31
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    • 2020
  • Five genes (mxbDM, mxcQE and mxaB) are responsible for the transcription of methanol oxidation genes in Methylobacterium strains. Among these, MxbDM and MxcQE constitute the two-component system (TCS) regulating methanol metabolism. In this study, we integrated the methanol-sensing domain of MxbD and MxcQ with the EnvZ/OmpR from Escherichia coli. The domain-swapping strategy resulted in chimeric histidine kinases (HK's) MxbDZ and MxcQZ AM1 containing recombinant E. coli. Real-time quantitative PCR was used to monitor OmpC expression mediated by the chimeric HK and response regulator (RR) OmpR. Further, an ompC promoter based fluorescent biosensor for sensing methanol was developed. GFP fluorescence was studied both qualitatively and quantitatively in response to environmental methanol. GFP measurement also confirmed ompC expression. Maximum fluorescence was observed at 0.05% methanol and 0.01% methanol using MxbDZ and MxcQZ AM1, respectively. Thus the chimeric HK containing E. coli were found to be highly sensitive to methanol, resulting in a rapid response making them an ideal sensor.

OmpR Is Essential for Growth and Expression of Virulence-related Genes in the Fish Pathogen Edwardsiella piscicida (어류 병원체 Edwardsiella piscicida의 OmpR은 생육과 병원성과 관련된 유전자의 발현에 필수적)

  • Ray, Durga;Kim, Yeon Ha;Choe, unjeong;Kang, Ho Young
    • Journal of Life Science
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    • v.31 no.1
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    • pp.28-36
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    • 2021
  • Edwardsiella piscicida is a significant cause of hemorrhagic septicemia in fish and gastrointestinal infections in humans. Survival bacteria require specialized mechanisms to adapt to environmental fluctuations. Hence, to understand the mechanism through which E. piscicida senses and responds to environmental osmolarity changes, we determined the protein expression profile and physiological properties under various salinity conditions in this study. The OmpR protein is a part of the Env-ZOmpR two-component system that has been implicated in sensing salt stress in bacteria. However, the physiological role played by this protein in E. piscicida remains to be elucidated. Therefore, in this work, the function of the OmpR protein in response to salt stress was investigated. Phenotypic analysis revealed that, in the mutant, three of the biochemical phenotypes were different from the wild type, including, citrate utilization, hydrogen sulfide, and indole production. Introduction of the plasmid containing the entire ompR gene to the mutant strain returned it to its parental phenotype. The retarded growth rate also partially recovered. Furthermore, in our studies, OmpR was not found to be related to cell motility. Taken together, our results from the mutational analysis, the growth assay, MALDI-TOF MS, qRT-PCR, and the phenotype studies suggest that the OmpR of E. piscicida is implicated in osmoregulation, growth, expression of porins (ETAE_1826), virulence-related genes (EseC, EseD and EvpC), and certain genes of unknown function (ETAE_1540 and ETAE_2706).

The Infectivity of Recombinant Porcine Endogenous Retrovirus (PERV-A/C) Is Modulated by Membrane-Proximal Cytoplasmic Domain of PERV-C Envelope Tail (C형 돼지 내인성 레트로바이러스(PERV)의 C-말단 외막당단백질에 의한 재조합 PERV-A/C의 감염력 조절)

  • Kim, Sae-Ro-Mi;Park, Sang-Min;Lee, Kyu-Jun;Lee, Yong-Jin;Bae, Eun-Hye;Park, Sung-Han;Lim, Ji-Hyun;Jung, Yong-Tae
    • Korean Journal of Microbiology
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    • v.46 no.1
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    • pp.15-20
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    • 2010
  • Xenotransplantation of pig organs is complicated by the existence of polytropic replication-competent porcine endogenous retroviruses (PERV) capable of infecting human cells. Two classes of infectious human-tropic replication-competent PERVs (PERV-A and PERV-B) and one class of ecotropic PERV-C are known. The potential for recombination between ecotropic PERV-C and human-tropic PERVs adds another level of infectious risk. A recombinant PERV-A/C (PERV-A14/220) virus is 500-fold more infectious than PERV-A. Two determinants of this high infectivity was identified; one was isoleucine-to-valine substitution at position 140 in RBD (receptor binding domain), and the other lies within the PRR (proline rich region) of the envelope protein. To examine whether the effects of the cytoplasmic tail of the PERV-C Env on fusogenesity also influences infectivity, we constructed a pseudotype retroviral vectors containing MoMLV core protein and PERV envelopes. Pseudotyping experiments with the PERV envelope glycoproteins indicated that recombinant PERV-A/C virus is 10-fold more infectious than PERV-A by lacZ staining. This result supports the suggestion that viral transduction of PERV-A/C is enhanced by a membrane-proximal cytoplasmic amphiphilic ${\alpha}$-helix in PERV-C Env tail.