• Title/Summary/Keyword: DnaJ

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Effect of Molecular Chaperones on the Soluble Expression of Alginate Lyase in E. coli

  • Shin, Eun-Jung;Park, So-Lim;Jeon, Sung-Jong;Lee, Jin-Woo;Kim, Young-Tae;Kim, Yeon-Hee;Nam, Soo-Wan
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.5
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    • pp.414-419
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    • 2006
  • When the alginate lyase gene (aly) from Pseudoalteromonas elyakovii was expressed in E. coli, most of the gene product was organized as aggregated insoluble particles known as inclusion bodies. To examine the effects of chaperones on soluble and nonaggregated form of alginate lyase in E. coli, we constructed plasm ids designed to permit the coexpression of aly and the DnaK/DnaJ/GrpE or GroEL/ES chaperones. The results indicate that coexpression of aly with the DnaK/DnaJ/GrpE chaperone together had a marked effect on the yield alginate lyase as a soluble and active form of the enzyme. It is speculated this result occurs through facilitation of the correct folding of the protein. The optimal concentration of L-arabinose required for the induction of the DnaK/DnaJ/GrpE chaperone was found to be 0.05mg/mL. An analysis of the protein bands on SDS-PAGE gel indicated that at least 37% of total alginate lyase was produced in the soluble fraction when the DnaK/DnaJ/GrpE chaperone was coexpressed.

Current Status and Development Direction of Advanced Air Mobility ICTs (Advanced Air Mobility ICT 기술 현황 및 발전 방향)

  • B.J. Oh;M.S. Lee;B.K. Kim;Y.J. Jeong;Y.J. Lim;C.D. Lim
    • Electronics and Telecommunications Trends
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    • v.38 no.3
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    • pp.1-10
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    • 2023
  • In this study, the status of global advanced air mobility (AAM) was investigated to derive information and communications technologies (ICTs) that should be prepared according to directions of domestic AAM development. AAM is an urban air traffic system for moving from city to city by electric vertical take-off and landing or personal aircraft. It is expected to establish a three-dimensional air traffic system that can solve ground traffic congestion caused by the rapid global urbanization. With the full-scale commercialization of AAM solutions, high-density air traffic is expected, and with the advent of the personal air vehicle (PAV), the flight space usage is expected to expand. Therefore, it is necessary to develop a safe AAM service through early research on core ICTs for autonomous flight.

Development of Restriction Fragment Length Polymorphism(RELP) Markers in Silkworm, Bombyx mori (누에 RFLP(제한단편 다형현상)마커 개발)

  • 고승주;김태산;이영승;황재삼;이상몽
    • Korean journal of applied entomology
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    • v.36 no.1
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    • pp.96-104
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    • 1997
  • A silkworm Bombyx mori genomic DNA library was constructed from polyphagous J111 strain and unpolyphagous $C_3$ strain to develop the genomic study by DNA makers. Genomic DNAs of two strains were digested with restriction enzyme EcoRI and ligated into pUC18. The ligated plasmids were transferred into E. coli host strain DH5$\alpha$. When the genomic DNAs were hybridized with insert DNAs from transformant, could be categorized from hybridization patterns to three groups as high repetitive sequence, moderately repetitive sequence, and low-copy number sequences. A total of 219 clones containing single or low-copy number sequence inserts were examined for any polymorphisms between two strains of J111 and $C_3$. Forty six clones showed RFLPs and 10 of these clones were used as a probe of analysis of $F_2$ population derived from crossing between J111 and $C_3$ strain. The genetic inheritance tested with each clones will be important tools to construct the genetic map of the silkworm, Bombyx mori.

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Studies on the Adenosine Deaminase Gene from Nocardioides sp. J-326TK (Nocardioides sp. J-326TK의 Adenosine Deaminase Gene에 관한 연구)

  • 전홍기;백형석;정춘식
    • Journal of Life Science
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    • v.8 no.6
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    • pp.673-680
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    • 1998
  • Adenosine deaminase gene from Nocardioides sp. J-326TK was cloned by polymerase chain reaction using primers (PI, PII and PIII) constructed from the highly conserved amino acid sequences among Escherichia coli, mouse and human. A PCR product of about 800bp, as expected from the sequence of E. coli adenosine deaminase gene, was obtained from Nocardioides sp. J-326TK chromosomal DNA double-digested with EcoRI and Pst I. DNA sequencing of the PCR product after cloning into pT7Blue T-vector shows 99.5% and 98.9% homologies in nucleotide and amino acid sequences, respectively, with the E. coli adenosine deaminase whereas 59.5% and 46.8% homologies with the human adenosine deaminase, indicating the evolutionarily relationship of these organisms.

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A Macrolide-Lincosamide-Streptogramin B Resistance Determinant Gene (ermJ) Cloned from B, anthracis 590

  • Kim, Hee-Sun;Choi, Eung-Chil;Kim, Byong-Kak;Park, Young-In
    • Archives of Pharmacal Research
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    • v.15 no.1
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    • pp.58-61
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    • 1992
  • Bacillus anthracis 590 having an inducibla resistance determinant to MLS antibiotics was isolated from a soli sample in Korea. The resistance gene (ernJ) was cloned by Southern blotting of chromosomal DNA fragment digested by various restriction enzymes and coloy hybridization method and the cloned plasmid was named as pBA423. The size of inserted DNA fragment of pBS42 vector was about 2.9 kb and the DNA sequence of the subcloned fragment (Hinc II-Hinc II, 1.4kb) WAS determined. The DNA sequence of ernJ was composed of 357 bp for leader region and 861 bp for the structural gene. Because the leader sequence of ernJ was homologous to that of ermK, the expression of ernJ is also thought to be controlled by a transcriptionl attenuation mechanism.

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Expression of Poly (ADP-ribose) Polymerase During Apoptosis Induced by Ultraviolet Radiation in HeLa $S_3$ Cells

  • Chang, Jeong-Hyun;Kwon, Heun-Young
    • Biomedical Science Letters
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    • v.12 no.4
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    • pp.349-354
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    • 2006
  • Induction of apoptosis allows the organism to get rid of abnormal cells and also of tumor cells. Understanding the mechanism involved in Ultraviolet radiation (UV) induced apoptosis may improve its therapeutic efficacy. In this study, we present expression of poly (ADP-ribose) polymerase (PARP) during apoptosis induced by UV in HeLa $S_3$ cells. Four different assays were performed in this study: morphological assessment of apoptotic cells and cell viability, DNA fragmentation analysis by agarose gel electrophoresis, quantitative assay of fragmented DNA, and expression of PARP by the western blot analysis. The percentages of apoptotic HeLa $S_3$ cells irradiated with $75J/m^2$ UV was increased continuously from 3 hrs incubation. DNA ladder pattern was appeared at 6 hrs. The amount of nucleosomal DNA fragments in cells treated UV increased from 3 to 12 hrs incubation and gradually decreased. The cleavage of PARP in HeLa $S_3$ cells irradiated with UV was induced, and the cleavage of PARP was more delayed in the cells pretreated with $5J/m^2$ UV and subsequently irradiated with $75J/m^2$ UV. than that in the cells only irradiated with $75J/m^2$ UV. Thus these data suggest that the cleavage of PARP relates with DNA fragmentation associated with apoptosis.

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Electrochemical Signal Detecting Method for DNA Sequencing (DNA 염기서열 분석을 위한 전기화학적 신호 검출 방법)

  • Cho, S.B.;Hong, J.S.;Yang, S.J.;Kwon, K.M.;Han, S.O.;Kim, Y.M.;Pak, J.H.
    • Proceedings of the KIEE Conference
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    • 2001.07c
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    • pp.1869-1871
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    • 2001
  • DNA 센서의 중요한 역할 중의 하나는 염기서열을 분석함으로써 유전적인 질병이나 돌연변이를 찾아낸다는 점이다. 염기서열 분석법으로 질량, 광학, 전기 화학적 측정법 등이 있는데, 그 중 전기 화학적 측정방법이 타 방법에 비해 간편하고 비용도 저렴해서 전망이 매우 밝다. 전기 화학적 측정을 위해서는 전극의 표면 처리 공정과 전극 표면에서의 DNA immobilization, hybridization 공정 및 전기적 신호를 발생시키는 intercalator, 그리고 전기적 신호 검출을 위한 측정 장비가 필요하다. 본 논문에서는 전극의 표면 처리 물질로서 2-mercaptoethanol을 사용했고 double strand DNA의 intercalator로써 methylene blue를 사용했으며, methylene blue의 환원 전류값을 측정하여 double strand DNA를 bare Au 또는 single strand DNA와 구분할 수 있었다. 이러한 연구 결과를 토대로 하여 전기 화학적 신호 검출을 이용한 DNA 센서의 가능성과 개발 방향을 제시하고자 한다.

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Genetic Relationship of Cordyceps spp. Based on Internal Transcribed Spacer Sequences of Ribosomal DNA (Ribosomal DNA의 ITS 염기서열에 의한 동충하초속균의 유연관계)

  • 남성희;황재삼;조세연;구태원
    • Journal of Sericultural and Entomological Science
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    • v.41 no.3
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    • pp.174-179
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    • 1999
  • The genetic relationships among six Cordyceps spp. were investigated based on internal transcribed spacer sequences of ribosomal DNA .A portion of the these genes was amplified by PCR. Approximately 590 base pairs were successfully amplified, cloned, sequenced, compared. The nucleotide sequence of the six amplified fragments were aligned by the clustal W program. As a result, Cordyceps militaris shared 87, 96, 98, 90 and 97% sequences homology with Paecilomyces japonica, Paecilomyces sp. J300, Paeciomyces farinosa. Paecilomyces sp. J500 and Cordyceps sinensis, respectively. Paecilomyces japonica also shared 87, 88, 92 and 87% sequence similarity with Paecilomyces sp. J300, Paecilomyces farinosa, Paecilomyces sp. J500 and Cordyceps sinensis, repectively.

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Differential Regulation of the Genes of the Streptococcus pneumoniae dnaK Operon by Ca++

  • Kim, Seung-Whan;Bae, Yong-Goo;Pyo, Suhk-Neung;Rhee, Dong-Kwon
    • Molecules and Cells
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    • v.23 no.2
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    • pp.239-245
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    • 2007
  • DnaK is a major antigen in Streptococcus pneumoniae, and is induced by a minor shift in temperature (30 to $37^{\circ}C$) but not by ethanol shock. Although HrcA in the presence of $Ca^{{+}{+}}$ represses the expression of both groEL and hrcA, the control of transcription of the dnaK operon is not completely understood. In this study, the dnaK operon of S. pneumoniae (5' hrcA-grpE-dnaK-dnaJ) was cloned and analyzed. It contains large intergenic regions in grpE/dnaK and dnaK/dnaJ. Pulse labeling with [$^{35}S$]-methionine and immunoblot analyses revealed the presence of higher levels of DnaK than of HrcA even in the presence of $Ca^{{+}{+}}$ after heat shock suggesting that $Ca^{{+}{+}}$ differentially regulates the heat shock responses of hrcA and dnaK. By blocking de novo mRNA synthesis with rifampin it was shown that neither the hrcA nor the groEL transcripts were stabilized by heat shock even though dnaK transcripts were stabilized. We conclude that S. pneumoniae uses fine regulation of the transcription of the individual genes of the tetracistronic dnaK operon to cope with the various stresses experienced during infections.

Low-dose of Ultraviolet radiation-, Ethyl methanesulfonateor Bleomycin-lnduced Adaptive Response in Chinese hamster ovary Cells

  • Lee, Dong-Wook;Shin, Eun-Joo;Um, Kyung-Il
    • Environmental Mutagens and Carcinogens
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    • v.15 no.2
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    • pp.94-99
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    • 1995
  • The adaptive response and cross-adaptive response to sister chromatid exchanges (SCEs) and DNA single-strand breaks (SSBs) in Chinese hamster ovary (CHO)-K$_1$ cells treated with ultraviolet radiation (UV), ethyl methanesuffonate (EMS), or bleomycin (BLM) were investigated. Two assays were used in this study; SCEs and alkaline elution. The pretreatment with low conditioning dose of 2 mM EMS or 1 J/m$^2$ UV decreased the yield of SCEs induced by subsequent treatment with 8 mM EMS, 5 J/m$^2$ UV or 5 $\mu$g/ml BLM. And the pretreatment with low conditioning dose of 1 $\mu$g/ml BLM decreased the yield of SCEs induced by subsequent treatment with 5 $\mu$g/ml BLM or 5 J/m$^2$ UV. The rejoining of DNA SSBs in cells subsequently treated with 2 J/m$^2$ UV, 50 mM EMS or 400 $\mu$g/ml BLM is higher than that only treated with 2 J/m$^2$ UV, 50 mM EMS or 400 $\mu$g/ml BLM. These results suggest that there are the adaptive response and cross-adaptive response to SCEs, and is the adaptive response to the rejoining of DNA SSBs in CHO cells.

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