• 제목/요약/키워드: chromosomal DNA

검색결과 520건 처리시간 0.028초

The Actin-Related Protein BAF53 Is Essential for Chromosomal Subdomain Integrity

  • Lee, Kiwon;Kim, Ji Hye;Kwon, Hyockman
    • Molecules and Cells
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    • 제38권9호
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    • pp.789-795
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    • 2015
  • A chromosome territory is composed of chromosomal subdomains. The internal structure of chromosomal subdomains provides a structural framework for many genomic activities such as replication and DNA repair, and thus is key to determining the basis of their mechanisms. However, the internal structure and regulating proteins of a chromosomal subdomain remains elusive. Previously, we showed that the chromosome territory expanded after BAF53 knockdown. Because the integrity of chromosomal subdomains is a deciding factor of the volume of a chromosome territory, we examined here the effect of BAF53 knockdown on chromosomal subdomains. We found that BAF53 knockdown led to the disintegration of histone H2B-GFP-visualized chromosomal subdomains and BrdU-labeled replication foci. In addition, the size of DNA loops measured by the maximum fluorescent halo technique increased and became irregular after BAF53 knockdown, indicating DNA loops were released from the residual nuclear structure. These data can be accounted for by the model that BAF53 is prerequisite for maintaining the structural integrity of chromosomal subdomains.

Binding of IciA protein to the dnaA promoter region

  • Kim, Hakjung;Hwang, Deog-Su
    • Journal of Microbiology
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    • 제33권3호
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    • pp.191-195
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    • 1995
  • IciA protein has been shown as an inhibitor for the initiation of E. coli chromosomal DNA replication at oriC. IciA protein binds the AT-rich region in oriC and then blocks the initiation of chromosomal DNA replication. Two binding sites for IciA protein were identified in dnaA gene, encoding the initiator for the E. coli chromosomal replication, promoter region by gel-shift assay and DNase I footprinting, One, named as IciA site I, is located upstream of the dnaA promoter 1P. The other, named as IciA site II, is located downstream of the dnaA promoter 2P. The sequence comparison of the regions protected from the DNase I cleavage did not result in a clear consensus sequence for the binding of IciA protein, suggesting that IciA protein may be a member of multimeric complex dsDNA binding proteins. This study provided information about the binding mode of IciA protein. Even though the IciA site II and IciA binding site in oriC seem to be composed of two IciA binding units, one binding unit is likely enough to cause the binding of IciA protein to the IciA site I. The binding of IciA protein to the dna4 promoter implies that IciA protein may involve not only the control of the initiation of chromosomal DNA replication but also the control of the dna4 gene expression.

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Genome-wide Examination of Chromosomal Aberrations in Neuroblastoma SH-SY5Y Cells by Array-based Comparative Genomic Hybridization

  • Do, Jin Hwan;Kim, In Su;Park, Tae-Kyu;Choi, Dong-Kug
    • Molecules and Cells
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    • 제24권1호
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    • pp.105-112
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    • 2007
  • Most neuroblastoma cells have chromosomal aberrations such as gains, losses, amplifications and deletions of DNA. Conventional approaches like fluorescence in situ hybridization (FISH) or metaphase comparative genomic hybridization (CGH) can detect chromosomal aberrations, but their resolution is low. In this study we used array-based comparative genomic hybridization to identify the chromosomal aberrations in human neuroblastoma SH-SY5Y cells. The DNA microarray consisting of 4000 bacterial artificial chromosome (BAC) clones was able to detect chromosomal regions with aberrations. The SH-SY5Y cells showed chromosomal gains in 1q12~ q44 (Chr1:142188905-246084832), 7 (over the whole chro-mosome), 2p25.3~p16.3 (Chr2:18179-47899074), and 17q 21.32~q25.3 (Chr17:42153031-78607159), while chromosomal losses detected were the distal deletion of 1p36.33 (Chr1:552910-563807), 14q21.1~q21.3 (Chr14:37666271-47282550), and 22q13.1~q13.2 (Chr22:36885764-4190 7123). Except for the gain in 17q21 and the loss in 1p36, the other regions of gain or loss in SH-SY5Y cells were newly identified.

Galangin 및 양강추출물의 $KBrO_3$ 유도 DNA 및 염색체 손상에 대한 보호효과 (Protective Effect of the 70% Ethanolic Extract of Alpinia officinarum and Galangin Against $KBrO_3$-induced DNA and Chromosomal Damage in Mice)

  • 양혜은;허문영
    • 약학회지
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    • 제54권6호
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    • pp.423-428
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    • 2010
  • The aim of this study was to evaluate the in vivo effect of galangin and the 70% ethanolic extract of Alpinia officinarum (AO) toward $KBrO_3$-induced DNA and chromosomal damage in mice. Galangin and AO inhibited the formation of 8-hydroxy-2'-deoxyguanosine (8-OH2'dG) as an indicator of DNA oxidative damage in the liver cell. Galangin and AO showed the inhibitory effect on the formation of DNA single strand break in the splenocyte by single cell gel electrophoresis (SCGE) assay and also inhibited micronucleated reticulocyte (MNRET) formation of peripheral blood in tail blood of mice. Vit-E revealed antigenotoxic effects in DNA and chromosome levels, but galangin was more potent active compound compare to vit-E under our experimental conditions. The results suggest that the extract of Alpinia officinarum containing galangin can modify the oxidative DNA and chromosomal damage and may act as chemopreventive agent against oxidative stress in vivo.

Serratia marcescens Metalloprotease 유전자의 대장균에로의 클로닝 (Molecular Cloning of Serratia rnarcescens Metalloprotease Gene into Escherichia coli)

  • 김기석;이창원;이상열;이병룡;신용철
    • 한국미생물·생명공학회지
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    • 제20권3호
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    • pp.280-288
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    • 1992
  • Serratia marcescens ATCC 21074 균주가 세포밖으로 분비하는 metalloprotease 유전자를 대장균으로 클로닝하고 그 발현을 살펴보았다 Serratia marcescens ATCC 21074 균주의 염색체 DNA를 제한효소 HindIII로 절단하고 아가로스 전기영동 후 32P로 표지된 합성 oligonucleotide를 사용하여 southern hybridization한 결과 4.0Kb의 DNA 절편에 metalloprotease가 존재함을 알 수 있었다. 4.0Kb 염색체 DNA 절ㅊ편을 분리하여 pUC19에 연결한 후 대장균으로 transformation하였다.

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염색체 말단부위 (Back to the Ends: Chromosomal DNA)

  • 이미형;서동철
    • Childhood Kidney Diseases
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    • 제12권1호
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    • pp.1-10
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    • 2008
  • Nucleic scids transfer the genetic information for serving a central biological purpose. The nucleic acids are polymers of nucleotides and they are mainly ribonucleic acid(RNA) and deoxyribonucleic acid(DNA). The nucleotides are stoichiometrically composed of five-carbon sugars, nitrogeneous bases, and phosphoric acids. The chemistry of nucleic acids and characteristics of different genomes are decribed for further study. Most of DNA genomes tend to be circular including bacterial genomes and eukaryotic mitochondrial DNA. Eukaryotic chromosomes in cells, in contrast, are generally linear. The ends of linear chromosomes are called telomeres. The genomes of different species, such as mammals, plants, invertebrates can be compared with the chromosome ends. The telomeric complex allows cells to distinguish the random DNA breaks and natural chromosomal ends. The very ends of chromosomes cannot be replicated by any ordinary mechanisms. The shortening of telomeric DNA templates in semiconservative replication is occurred with each cell division. The short telomere length is critically related to aging, tumors and dieases.

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NCAPH Stabilizes GEN1 in Chromatin to Resolve Ultra-Fine DNA Bridges and Maintain Chromosome Stability

  • Kim, Jae Hyeong;Youn, Yuna;Hwang, Jin-Hyeok
    • Molecules and Cells
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    • 제45권11호
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    • pp.792-805
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    • 2022
  • Repairing damaged DNA and removing all physical connections between sister chromosomes is important to ensure proper chromosomal segregation by contributing to chromosomal stability. Here, we show that the depletion of non-SMC condensin I complex subunit H (NCAPH) exacerbates chromosome segregation errors and cytokinesis failure owing to sister-chromatid intertwinement, which is distinct from the ultra-fine DNA bridges induced by DNA inter-strand crosslinks (DNA-ICLs). Importantly, we identified an interaction between NCAPH and GEN1 in the chromatin involving binding at the N-terminus of NCAPH. DNA-ICL activation, using ICL-inducing agents, increased the expression and interaction between NCAPH and GEN1 in the soluble nuclear and chromatin, indicating that the NCAPH-GEN1 interaction participates in repairing DNA damage. Moreover, NCAPH stabilizes GEN1 within chromatin at the G2/M-phase and is associated with DNA-ICL-induced damage repair. Therefore, NCAPH resolves DNA-ICL-induced ultra-fine DNA bridges by stabilizing GEN1 and ensures proper chromosome separation and chromosome structural stability.

Agrobacterium tumefaciens의 염색체 DNA내에 존재하는 종양 유발 지역의 클로닝 (Cloning of agrobacterium tumefaciens chromosomal virulence region)

  • 심웅섭
    • 미생물학회지
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    • 제28권2호
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    • pp.104-108
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    • 1990
  • Ti-plasmid에 의하여 식물체에 종양이 형성되기 위해서는 맨먼저 Agrobacteriuin tumefariens가 식물세포에 부착되어야 하는데, 이에 필요한 유전자들은 세균의 염색체 DNA내에 존재한다. 이들 유전자를 클로닝하기 위하여 transposon 쑤5를 사용하여 A. tumefaciens A5512에 돌연변이를 유발시켰다. 그 결과 식물세포에 부착하지 못하며, 종양을 유발시키지 못하는 두개의 돌연변이체를 얻었다. 이 두 돌연변이체 중 하나로부터 세균이 식물세포에 부착하는데 관여하는 염색체 DNA내의 종양 유발 지역을 클로닝하였다.

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Polymerase Chain Reaction에 의한 Lactobacillus casei 및 돌연변이 균주들의 비교 분석 (Analysis of Lactobacillus casei and Mutant Strains by Polymerase Chain Reaction)

  • 남진식;이정준;신명수;나석환;백영진;유민
    • 한국미생물·생명공학회지
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    • 제22권6호
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    • pp.577-583
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    • 1994
  • To classify Lactobacillus casei strains on the basis of difference in their chromosomal DNA sequence, we have performed polymerase chain reactions on their chromosomal DNA by using random primers, and followed by analyzing randomly amplified polymorphic DNA fragments. We also developed a mini-preparative method to isolate PCR-grade chromosomal DNA from Lacto- bacillus casei strains within 3 hours. Based on RAPD pattems by polymerase chain reactions with degenerated random primers, 4 Lactobacillus casei strains and 2 mutant strains were successfully discriminated. Results were very sensitive, strain-specific and reproducible. It was also reliable. These results suggest that RAPD may be applied efficiently for the identification of several Lactoba- cillus casei strains.

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Yarrowia lipolytica의 Multicopy Integration Vector 개발 (Development of a Multicopy Integration Vector in Yarrowia lipolytica)

  • 김정윤;우문희
    • 한국미생물·생명공학회지
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    • 제23권5호
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    • pp.536-543
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    • 1995
  • Multicopy integration vector is a very useful vector system in that they can be integrated into chromosomal DNA in several copies and stably maintained under non-selective conditions. To develop a multicopy integration vector system in the yeast Yarrowia lipolytica, P-type ribosomal DNA was cloned from Y lipolytica. A HindIII-BglII fragment of the cloned rDNA and a promoterless URA3 gene were inserted into pGEM1, generating multicopy integration vectors, pMIYL-1 and pMIYL-2. The rDNA fragment is for targeted homologous recombination between the vector and the chromosomal DNA of Y. lipolytica, and the promoterless URA3 gene is a defective selection marker for inducing multicopy integration. pMIYL-1 and pMIYL-2 have an unique restriction enzyme site, KpnI, and two unique restriction enzyme sites, KpnI and EcoRI, repectively, which can be used for targeting of the vectors into the rDNA of Y. lipolytica chromosomal DNA. After transformation of the vectors into Y. lipolytica, copy number and stability were analyzed by Southern hybridization. The vectors were found to be present in less than 5 copies per cell and were stably maintained during growth in non-selective media.

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