• Title/Summary/Keyword: DNA.

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Genome Detection Using an DNA Chip Array and Non-labeling DNA (비수식화 바이오칩 및 유전자 검출)

  • Choi, Yong-Sung;Lee, Kyung-Sup
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2006.06a
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    • pp.402-403
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    • 2006
  • This research aims to develop the multiple channel electrochemical DNA chip using microfabrication technology. At first, we fabricated a high integration type DNA chip array by lithography technology. Several probe DNAs consisting of thiol group at their 5-end were immobilized on the gold electrodes. Then target DNAs were hybridized and reacted. Cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. Therefore, it is able to detect a plural genes electrochemically after immobilization of a plural probe DNA and hybridization of non-labeling target DNA on the electrodes simultaneously. It suggested that this DNA chip could recognize the sequence specific genes.

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SNP Detection Using Indicator-free DNA Chip (비수식화 DNA를 이용한 유전자 검출)

  • Choi, Yong-Sung;Moon, Jong-Dae;Lee, Kyung-Sup
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2006.06a
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    • pp.410-411
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    • 2006
  • High throughput analysis using a DNA chip microarray is powerful tool in the post genome era. Less labor-intensive and lower cost-performance is required. Thus, this paper aims to develop the multi-channel type label-free DNA chip and detect SNP (Single nucleotide polymorphisms). At first, we fabricated a high integrated type DNA chip array by lithography technology. Various probe DNAs were immobilized on the microelectrode array. We succeeded to discriminate of DNA hybridization between target DNA and mismatched DNA on microarray after immobilization of a various probe DNA and hybridization of label-free target DNA on. the electrodes simultaneously. This method is based on redox of an electrochemical ligand.

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NMR Study of Temperature-Dependent Single-Stranded DNA Binding Affinity of Human Replication Protein A

  • Kim, Min-Gyu;Shin, Tae-Hoan;Choi, Seo-Ree;Choi, Jae-Gyu;Lee, Joon-Hwa
    • Journal of the Korean Magnetic Resonance Society
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    • v.20 no.3
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    • pp.66-70
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    • 2016
  • The replication protein A (RPA), is a heterotrimer with 70, 32 and 14 kDa subunits and plays a crucial role in DNA replication, recombination, and repair. The largest subunit, RPA70, binds to single-stranded DNA (ssDNA) and mediates interactions 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 temperatures, to understand the temperature dependency of ssDNA binding affinity of RPA70A. Essential residues for ssDNA binding were conserved while less essential parts were changed with the temperature. Our results provide valuable insights into the molecular mechanism of the ssDNA binding of human RPA.

Synthesis and Characterization of DNA-mediated Gold Nanoparticles by Chemical Reduction Method (화학적환원에 의한 DNA-mediated 금 나노입자의 합성 및 특성)

  • Sohn, Jun Youn;Sohn, Jeong Sun
    • Applied Chemistry for Engineering
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    • v.26 no.4
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    • pp.515-519
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    • 2015
  • Complexes composed of hydrogen tetrachloroaurate (III) trihydrate ($HAuCl_4{\cdot}3H_2O$) and DNA were first formed for the synthesis of gold nanoparticle using a DNA template, which were validated using UV-Vis spectroscopy. The morphology of complexes were also characterized by scanning electron microscopy (SEM). DNA-mediated gold nanoparticles were synthesized by the chemical reduction of DNA-Au(III) complexes using hydrazine ($N_2H_4$) and sodium borohydride ($NaBH_4$) as reducing agents. The effects of reducing agent types and their concentration on the formation of gold nanoparticles were investigated. The results showed that hydarazine was the most effective for the reduction of DNA-Au(III) complex. The DNA-mediated gold nanoparticles were characterized SEM, particle size analyzer (PSA), and transmission electron microscopy (TEM). Gold nanoparticles with 55~80 nm in diameter were formed by the aggregation of smaller gold nanoparticles (~nm), which was confirmed in the DNA matrix.

DnaJC18, a Novel Type III DnaJ Family Protein, is Expressed Specifically in Rat Male Germ Cells

  • Gomes, Cynthia;Soh, Jaemog
    • Development and Reproduction
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    • v.21 no.3
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    • pp.237-247
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    • 2017
  • Mammalian spermatogenesis occurs in a precise and coordinated manner in the seminiferous tubules. One of the attempts to understand the detailed biological process during mammalian spermatogenesis at the molecular level has been to identify the testis specific genes followed by study of the testicular expression pattern of the genes. From the subtracted cDNA library of rat testis prepared using representational difference analysis (RDA) method, a complimentary DNA clone encoding type III member of a DnaJ family protein, DnaJC18, was cloned (GenBank Accession No. DQ158861). The full-length DnaJC18 cDNA has the longest open reading frame of 357 amino acids. Tissue and developmental Northern blot analysis revealed that the DnaJC18 gene was expressed specifically in testis and began to express from postnatal week 4 testis, respectively. In situ hybridization studies showed that DnaJC18 mRNA was expressed only during the maturation stages of late pachytene, round and elongated spermatids of adult rat testis. Western blot analysis with DnaJC18 antibody revealed that 41.2 kDa DnaJC18 protein was detected only in adult testis. Immunohistochemistry study further confirmed that DnaJC18 protein, was expressed in developing germ cells and the result was in concert with the in situ hybridization result. Confocal microscopy with GFP tagged DnaJC18 protein revealed that it was localized in the cytoplasm of cells. Taken together, these results suggested that testis specific DnaJC18, a member of the type III DnaJ protein family, might play a role during germ cell maturation in adult rat testis.

Comparison of the Efficiency from Raw and Processed Corns by Five Different DNA Extraction Methods (다섯 가지 DNA 추출방법에 의한 옥수수 원료 및 가공시료의 DNA 추출 효율의 비교)

  • Lee, Hun-Hee;Song, Hee-Sung;Kim, Jae-Hwan;Lee, Woo-Young;Lee, Soon-Ho;Park, Sun-Hee;Park, Hye-Kyung;Kim, Hae-Yeong
    • Applied Biological Chemistry
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    • v.48 no.4
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    • pp.331-334
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    • 2005
  • In this study, the effects of five extraction methods for raw and processed corns were compared with respect to the integrity, yields and quality of DNA extracted from them and the results were assessed by PCR analysis. From the comparison of five extraction methods, DNA integrity showed a similar pattern. Amounts of genomic DNA obtained from the five extraction methods varies from $0.25{\mu}g\;to\;234{\mu}g$ per 1 g sample. The DNA yield extracted with CTAB method and DNeasy Plant Maxi kit is greater than that obtained from other extraction methods. These results would be applicable for the selection of an adequate extraction method for specific samples.

Expression of Canavalia Iineata Leghemoglobin cDNA in Transgenic Nicotiana tabacum (형질전환된 담배에서 해녀콩 Leghemoglobin cDNA의 발현)

  • 이선영
    • Journal of Plant Biology
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    • v.38 no.2
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    • pp.203-209
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    • 1995
  • Tobacco (Nicotiana tahacum L. cv. Wisconsin 38) leaf discs were cocultivated with Agrohacterium carrying a leghemoglobin (Lb) cDNA from Canavalia lineata. Seven plants were regenerated from the transformed leaf discs on MS media supplemented with 0.5 mg/L BAP, 0.1 mg/L ${\alpha}-NAA$, 200 mg/L kanamycin and 500 mg/L carbenicillin. Southern hybridization and PCR of genomic DNA from transgenic plants showed that the Lb cDNA was stably integrated into the genome of the tobacco. Total RNA from the transgenic tobacco showed northern hybridization signal at 1,000 nt and PCR of the first strand cDNA synthesized from the total RNA amplified 0.5 kb Lb cDNA. Furthermore, western hybridization using a polyclonal antibody against soybean Lb showed a 15.8 kD LB-like band on SDS-PAGE of proteins from the transformed tobacco. These results demonstrated that the Lb cDNA of C. lineata was not only incorporated into the genome of tobacco, but also transcribed into mRNA and translated into Lb protein in the transformed tabacco.

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Construction of the cDNA Library from Bombyx mori Larvae and Analysis of the Partial cDNA Sequences (누에 유충의 cDNA 유전자 은행 제작 및 cDNA 클론의 부분염기서울 분석)

  • 김상현;윤은영
    • Journal of Sericultural and Entomological Science
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    • v.38 no.1
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    • pp.13-18
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    • 1996
  • To secure the genetic resources of silkworm, Bomyx mori, the cDNA library was constructed with mRNA isolated from fifth instar larvae. Titer of the cDNA library was about 1.3 X 106 plaques in total. We presumed that the titer covered all transcripts existed in Bombyx mori. Meanwhile, it is knowen that partial cDNA sequences, Expressed Sequence Tags(ESTs), have a good value for the discovery of novel genes and the elucidation of their structures. For this purpose, partial cDNA sequencing was carried out from randomly selected cDNA clones in the library. Partial cDNA sequences of 37 clones were determined and an average of 212 nucleotides of sequence can be read from the clone. The ESTs were searched in GenBAnk database and fifteen ESTs showed significant similarities to enlisted sequences. They included the genes of storage protein, heat shock protein, actin, catalase and so forth. We presumed that the 22 unmatched ESTs were novel genes.

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A study for a construction of Theileria sergenti DNA probe (Theileria sergenti DAN probe를 만들기 위한 기초 연구)

  • Kim, Myeong-chul;Lee, Joo-mook;Kwon, Oh-deog;Chae, Joon-seok;Kim, Heung-seob
    • Korean Journal of Veterinary Research
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    • v.33 no.3
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    • pp.479-486
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    • 1993
  • This study was attempted co develope a method for detection of Theileria sergenti infection on the basis of hybridization of parasite DNA with a probe. For construction of a T sergenti genomic library, T sergenti DNA was digested completely with Bam-HI and the fragments were ligated into the Bam-HI site of pUC-19 before transformation of Escherichia colistrain JM83. To detect clones containing the parasite's DNA sequences, a genomic DNA library of T sergenti constructed in pUC-19 was screened by cracking and Southern hybridization. Seven colonies were chosen from 29 colonies which were screened by transformation of Escherichia coli strain JM83. Seven transformants were comfirmed from seven colonies by cracking. The sizes of transformants were about 5Kb, 5.7Kb, 4.3Kb, 7.75Kb, 7.85Kb, 5.8Kb, 3.8Kb, respectively. DNA inserts, T sergenti DNA, and bovine DNA were hybridized with radio-labelled T sergenti DNA. Two($pT_1$, $pT_1$) of the seven inserts and T sergenti DNA reacted strongly but another 5 inserts and bovine DNA showed weak reation. All of the DNA inserts were not reaction, but T sergenti DNA were very weakly and bovine DNA were strongly reacted to hybridization with radio-labelled bovine DNA. Therefore, we obtained total 7 T sergenti DNA fragments in this study.

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Directed Alignment of DNA Molecule between the gold electrodes (금 전극위에 DNA 분자의 정렬에 관한 연구)

  • Hwang, Hyun Suk;Kim, Hyung Jin
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.8
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    • pp.5586-5590
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    • 2015
  • In this paper, the directed alignment methode of the DNA molecule between the Au electrodes was suggested for the application of nano devices. To fabricate the nano device coated DNA, 2-Aminoethanthiol(AET) was coated on Au electrodes which was formed using photo-lithography process on $SiO_2/Si$ substrates. In general, the AET that was a positive charge with $NH^{3+}$ was strongly combined under the electrostatic interaction with DNA molecule which had to be a negative charge. The DNA molecules could be easily aligned between Au electrodes coated with AET. The structures of the DNA molecules were investigated using AFM(Atomic force microscope), they were changed from single types to bundle according to the AET concentrations.