• Title/Summary/Keyword: $P_{dnaK}$

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Preparation and In Vitro Release of DNA-Loaded Poly(D,L-lactic-co-glycolic acid) Microspheres (DNA가 봉입된 Poly(D,L-lactic-co-glycolic acid) 미립구의 제조 및 시험관내 방출)

  • Son, Hye-Jung;Kim, Jin-Seok
    • Polymer(Korea)
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    • v.29 no.1
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    • pp.69-73
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    • 2005
  • To overcome the main disadvantages of non-viral gene delivery systems such as repeated administration due to the low transfection efficiency, poly(D,L-lactide-co-glycolide) was applied to encapsulate pDNA in its microsphere formulation. Free pDNA or various ratios (w/w) of chitosan/pDNA complexes was used for encapsulation, with the resulting encapsulation efficiency of 44%, 5%, and 8% for free pDNA, 0.7:1 and 1:1 ratios, respectively. Scanning electron micrographs of poly(D,L-lactic-co-glycolic acid) (PLGA) microspheres encapsulating pDNA or chitosan-condensed pDNA revealed a smooth spherical shape immediately after microsphere preparation and a collapsed porous shape in 41 days due to the degradation of PLGA. In vitro release profile showed that the 0.7:1 (w/w) ratio formulation exerted 47% release in 26 days, whereas free pDNA or 1:1 (w/w) ratio formulation did only 15% or 32%, respectively.

NMR Study of the pH Effect on the DNA Binding Affinity of Human RPA

  • Lee, Min-Woo;Choi, Ju-Hyeok;Choi, Jae-Gyu;Lee, Ae-Ree;Lee, Joon-Hwa
    • Journal of the Korean Magnetic Resonance Society
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    • v.20 no.3
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    • pp.71-75
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    • 2016
  • The replication protein A (RPA) plays a crucial role in DNA replication, recombination, and repair. RPA consists of 70, 32 and 14 kDa subunits and has high single-stranded DNA (ssDNA) binding affinity. The largest subunit, RPA70, mainly contributes to bind to ssDNA as well as interact with many cellular and viral proteins. In this study, we performed nuclear magnetic resonance experiments on the complex of the DNA binding domain A of human RPA70 (RPA70A) with ssDNA, d(CCCCC), at various pH, to understand the effect of pH on the ssDNA binding of RPA70A. The chemical shift perturbations of binding residues were most significant at pH 6.5 and they reduced with pH increment. This study provides valuable insights into the molecular mechanism of the ssDNA binding of human RPA.

Molecular Cloning and Expression of Human Poly (ADP-ribose) Synthetase cDNA in E. Coli (인간 Poly(ADP-ribose) Synthetase cDNA의 클로닝 및 대장균에서의 발현)

  • 이성용;김완주;이태성;박상대;이정섭;박종군
    • The Korean Journal of Zoology
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    • v.39 no.3
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    • pp.248-256
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    • 1996
  • The present study was performed to clone and express human poly (ADP-ribose) synthetase (PARS) cDNA in E coli. For these purposes, the CDNA for human poly (ADP-ribose) synthetase, encoding the entire protein, was cloned into pGEM-7Zf(+). The resulting recombinant plasmid pPARS6.1 was restriction enzyme mapped and its identity was confirmed by Southern blot analysis. The pPARS6. 1 contained full-length CDNA of human PARS and the nudeotide sequences were identical with those reported previously. The recombinant protein which migrated as a unique 120 kDa band on 10% SDS-polyacrylamide gels, was identified as PARS by Southwestern blots using nick-translated DNA probes and by activity gels and activity blots using 32 P-NAD as a substrate for poly (ADP-ribose) synthetase (PARS). The signals corresponding to 120 and 98 kDa proteins were obtained following IPTG (0.4 mM) induction of the PARS cDNA cloned into Xba I-digested pGEM-7Zf(+) vector. Nonspecific signals corresponding to 45 and 38 kDa proteins were also shown in both IPTG-induced and noninduced cells. The nonspecific proteins may be products of incomplete translation or proteolytic products of intact PARS.

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Study on isolation of Prevotella nigrescens 9336- specific DNA probes using random cloning method (무작위 클로닝법을 이용한 Prevotella nigrescens 9336 특이 DNA 프로브의 개발에 관한 연구)

  • Gang, Soon-Won;Kim, Se-Hoon;kim, Dong- Ki;Seong, Jin-Hyo;Kim, Byung-Ock;Kim, Jung- Ki
    • Journal of Periodontal and Implant Science
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    • v.32 no.2
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    • pp.269-280
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    • 2002
  • The purpose of this study is to develop species-specific DNA probes and polymerase chain reaction (PCR) primers for detection and identification of Prevotella nigrescens (P. nigrescens) 9336. This study procedure includes (1) whole-genomic DNA extraction of P. nigrescens 9336 (2) construction of the genomic DNA library, (3) screening of strain-specific DNA probe by reverse Dot Hybridization method, (4) confirmation of strain-specific DNA probe by Southern blot analysis, (5) determination of nucleotide sequences of strain-specific DNA probe. Thirty-five restriction fragments of P. nigrescens 9336 genomic DNA digested with the Hind III were obtained. Reverse dot hybridization and Southern blot analysis data showed that three of them, Pn10, Pn23, and Pn35, could be P. nigrescens 9336-specific DNA probes. These data indicated that these DNA probes could be useful in detection and identification of the P. nigrescens 9336.

Effect of pH on Synthesis of Polypyrrole Nanowires by Using DNA Molecule Templates (DNA 분자를 형틀로 이용한 Polypyrrole 나노와이어의 합성시 pH 효과)

  • Choi, Young-Hun;Kim, Kyoung-Soeb;Kim, Nam-Hoon;Roh, Yong-Han
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2007.11a
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    • pp.109-110
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    • 2007
  • Pyrrole and DNA can be used for synthesis of conducting nanowires. Protonated pyrrole and negatively charged DNA are absorbed by electrostatic interaction. The level of absorbance is related to pH of pyrrole. Therefore, DNA immobilized and aligned on the 3-aminopropyltrimethoxysi1ane (APTES) modified Si surface. Positive pyrrole monomers was deposited on aligned DNA for the synthesis of nanowire in various pH condition. And polypyrrole nanowires were synthesized by polimerization with ammonium persulfate (APS). These polypyrrole nanowires were measured by AFM, and then we found optimal pH level for the synthesis of nanowires.

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Development of Repetitive DNA Probes for Genetic Analysis of Phytophthora capsici (Phytophthora capsici의 유전적 특성 분석을 위한 Repetitive DNA Probe의 개발)

  • Song, Jeong-Young;Kim, Hong-Gi
    • The Korean Journal of Mycology
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    • v.30 no.1
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    • pp.66-72
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    • 2002
  • To develop DNA markers for analysis of genetic characteristics of Phytophthora capsici population, randomly selected clones from HindIII-digested genomic DNA library of P. capsici 95CY3119 were surveyed by hybridizing to Southern blots of HindIII-digested total genomic DNA of P. capsici. Probe DNAs inserted into selected individual clones strongly hybridized with HindIII digests of P. capsici. Among probes examined, PC9 revealed the repetitive and highly polymorphic bands to HindIII digests of inter-and intra-field P. capsici isolates. Genetic diversity of individual isolates was also clearly revealed in cluster analysis based on its band patterns. The other probe, PC22, was hybridized only to DNA from P. capsici and this was highly repetitive. However, there was no response to other Phytophthora species and Pythium sp. These DNA probes could be used as very useful markers in analysing genetic diversity and identification for P. capsici population throughout the world.

Polyplex Formation of Calf Thymus DNA with Branched and Linear Polyethyleneimine

  • Han, Jung-Ho;Kim, Seog K.;Cho, Tae-Sub;Lee, Jae-Cheol;Joung, Hyun-Sook
    • Macromolecular Research
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    • v.12 no.5
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    • pp.501-506
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    • 2004
  • We have investigated the spectral properties of DNA, including its electric absorption, circular and linear dichroism (CD and LD), and fluorescence emission, in the DNA-linear polyethyleneimine (LPEI) and DNA-branched polyethyleneimine (BPEI) complexes at various polymer concentrations. The spectral properties of both complexes are similar. We observed a relatively moderate change in the absorption and CD spectra at low amine/DNA phosphate (NIP) ratios (< 0.5), followed by a drastic collapse within the N/P range from 0.8 and 1.0. The absorption and CD spectra recovered as the N/P ratio increased to ca. 1.2. In contrast, the LD and emission of ethidium intercalated between the DNA bases decreased almost linearly at N/P ratios between 0.0 and 1.0. These spectra never recovered at higher N/P ratios. We believe that the moderate changes in the spectrum at low N/P ratios occurred because of electrostatic interactions between DNA and BPEI, while the collapsed spectra at N/P ratios between 0.5 and 1.5 occurred because of condensation/aggregation of the DNA. Considering the structure of the polymers, we suggest that the secondary amino group of LPEI and all three amino groups of BPEI are equally involved in DNA condensation.

Analysis of DNA Conformation in the Particles of Bacteriophage P4 Mutant, P4 ash8 (박테리오파아지 P4 ash8 sid71 입자 내 DNA 형태 분석)

  • Song, Jae-Ho;Kim, Kyoung-Jin
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.62-66
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    • 2006
  • To study the packaging mechanism of the bacteriophage P2-P4 system which is a useful experimental tool for the study of viral capsid assembly, we analyzed the DNA contents of P4 sid- mutant, P4 ash8 sid71's phage particles. Two kind of particles having different density were separated by the CsCl buoyant equilibrium density gradient experiment with fresh made stock of P4 ash8 sid71. The DNA from each particles was prepared and its conformations was analyzed by electrophoresis. Unexpectedly, both particles contain not only dimeric and trimeric but also monomeric P4 DNA.

cis-Diamminedichloroplatinum (II) induces denaturation and conformational changes in pBR322 DNA (cis-Diamminedichloroplatinum(II)에 의한 pBR322 DNA의 변성과 구조 변화)

  • Koo, Ja-Choon;Lim, Chang-Soo;Hahn, Tae-Ryong;Yang, Jai-Myung
    • Applied Biological Chemistry
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    • v.33 no.4
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    • pp.343-348
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    • 1990
  • E. coli LE392, transformed with CDDP-treated pBR322 DNA, was plated on ampicillin containing media. The number of colonies formed on ampicillin containing agar plate was reduced to undetectable level after treat the DNA with 13.3 ${\mu}M$ CDDP. The CDDP-treated pBR322 DNA was susceptible to sing1e strand DNA specific S1 nuclease and it's migration Pattern in agarose gel electrophoresis was changed. These results suggest that CDDP adduction to pBR322 DNA resulted in denaturation of the double helix and changes in it's conformation which ultimately leads In the inactivation of the ampicillin resistant sere.

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Genetic Variation and Phylogenetic Relationship of Korean Ginseng based on cpDNA trnL-F, nrDNA ITS and ETS Sequences (엽록체 DNA trnL-F 및 핵리보조옴 DNA ITS, ETS 염기서열에 의한 고려인삼의 유전적 변이와 계통학적 유연관계)

  • Bang Chan Kuk;Kim Ju Hwan;Baek Myeong Hyun;Kim Chang Sik;Um Dong Myeong;Kim Dong Hee
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.6
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    • pp.1699-1709
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    • 2004
  • Genetic variations and relationship based on the sequences of cpDNA trnL-F gene, nrDNA ITS and ETS region among the twenty four taxa including Panax ginseng C.A. Meyer and its related species were investigated. And taxonomic status and molecular phylogenetic relationship between P. ginseng and related groups were discussed. Molecular systematic data from cpDNA and nrDNA sequences were very useful to elucidate the genetic variations and relationships among the treated taxa. It was found that P. ginseng is the independent unique species with distinct genetic limitation from the related species such as P. quinquefolius, P. japonicum, P. notoginseng and P. pseudoginseng. P. ginseng including cultivated types as well as wild ones formed monophyletic group with high genetic similarities. P. quinquefolius and P. japonicum were the most related sister groups of P. ginseng based on the molecular phylogenetic results in this study.