• Bulletin of the Korean Chemical Society
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• v.21 no.10
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• pp.1025-1030
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• 2000

• Bulletin of the Korean Chemical Society
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• v.21 no.6
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• pp.551-552
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• 2000

• Journal of Life Science
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• v.19 no.3
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• pp.305-310
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• 2009

Plasmid pGKV21 contains a 229-nucleotide (nt) single-strand DNA initiation (ssi) signal. Using asymmetric PCR, we prepared a small single-stranded (ss) DNA fragment of the ssi signal and, using the 229-nt ssDNA fragment, determined the requirements of RNA polymerase for priming and DNA-protein interaction. The ssi fragment prepared was able to generate primer RNAs with almost the same efficiency as the $M13{\Delta}lac182/229$ phage DNA. However, the cssi (complementary strand of the ssi signal) fragment could not synthesize primer RNAs. This result suggests that the 229-nt ssi signal functions in a strand specific manner. Gel retardation and DNase I footprinting demonstrated that the synthesized ssi fragment could interact with both E. coli RNA polymerase and SSB protein to synthesize primer RNA. In Escherichia coli [pWVAp], an addition of rifampicin resulted in an accumulation of ssDNA, indicating that the host-encoded RNA polymerase is involved in the conversion of ssDNA to double-stranded plasmid DNA.

• Korean Journal of Veterinary Research
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• v.15 no.1
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• pp.57-67
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• 1975

The poxvirus group is considered to be a typical cytoplasmic inclusion forming virus. Every poxvirus has been reported to produce only one kind of inclusion in the infected tissues. A vague concept that inclusions of poxviruses are eosinophilic or acidophilic has prevailed. Although many papers and theories about the nature of the inclusion have been presented, most of them are not quite convincing on the point of the relations with virus multiplication, and an analysis of papers published showed that there seem to be many discrepancies in the descriptions of the nature of the poxvirus inclusions. Comparative studies on host-virus interaction with cowpox, orf, swinepox and fowlpox viruses which selected from each Group (I-IV) of poxviruses were performed from the morphological and virological standpoints. At first, in cowpox virus-FL cell system, as a comparative model, cytoplasmic inclusion, nucleic acid metabolism by autoradiography and detection of viral antigen by immunofluorescence were studied and obtained the results as follows: 1. The focus-like cytopathic effect (CPE) at early stage developed to entire culture at terminal stage of infection, and also the developing status of CPE was correlated to viral doses for inoculation. Two kinds of cytoplasmic inclusions which named A and B type were easily observed by Giemsa, hematoxylin-eosin (H & E) and May-Greenwald Giemsa (MGG) stainings in the infected cells. The B type inclusions were formed at early stage of infection and the A type inclusions were produced subsequently the B type formation. The B type which common type inclusion in poxviruses was a small compact or aggregate at early stage and developed to a large diffuse body at terminal stage of infection. On the other hand, the A type inclusion which depend upon the kind of virus was appeared as round and discrete shape, and its size and number was increased gradually during the culture period. It was characteristic to form distinct halos around the both types of inclusions in acid fixed, H & E stained preparations of infected cultures. The B type inclusion was always positive in Feulgen reaction and showed as DNA containing body but the A type inclusion was not. 2. In the relationship between inclusion and DNA metabolism of infected cells by the qualitative autoradiography using 3H-thymidine, the appearance of silver grains was coincided with B type inclusion but not with A type inclusion. This showed that the DNA synthesis was proceeded in all B type inclusions except those in the terminal stage with a diffuse form. This suggested that the B type inclusions are only sites of DNA synthesis and this was proceeded after the cell infection independently. The activity of DNA synthesis of the inclusions was nearly the same as that of the nucleic of normal cells and non-inclusion bearing cells. and non-inclusion bearing cells. Regardless of the size of the degree of DNA synthesis of the B type inclusion, inclusion bearing cells all showed remarkable suppression of nuclear DNA synthesis. 3. By the direct fluorescent antibody technique viral antigen in infected cells was detected. The B type inclusions have been proved to contain a great deal of viral antigen, whereas the basic substance of A type inclusion did not show antigenicity except the round edge. It was suggested that the round edge fluorescence might be caused by the glare of cytoplasmic viral antigen which pushed out and concentrated by the A type inclusion development. 4. Hemorrhagic red pock formations on chorioallantoic membrane of embryonated chicken egg had proved the characteristic of used viral strain. 5. By the above studies on the nature of two types of inclusions and the role they play in virus multiplication, it was concluded that the B type inclusion must be the site of the synthesis of viral DNA and protein as well as the site of the virus.

• Proceedings of the Korea Crystallographic Association Conference
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• 2003.05a
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• pp.16-16
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• 2003

Mismatches in a DNA duplex are mainly due to DNA duplication errors that are generated by improper function of DNA polymerase. MutS, MutL and MutH are crucial proteins for the initiation of the methyl-directed mismatch repairing in bacteria. MutS has an ATPase activity md recognize the mismatched or unpaired bases on DNA. After binding to a mismatch, MutS recruits MutL to mediate the activation of MutH an endonuclease, which cleaves the 5' site of d(GATC) on the un-methylated strand. Both MutL and MutS also have essential roles in the subsequent removal and re-synthesis of the daughter strand. We have determined the crystal structures of either intact or active fragments of each of these proteins, both alone and complexed with ligands (DNA, ADP and ATP). The biochemical and mutagenesis studies based on the detailed 3-D structures led to new insights into the role of the ATPase activity of MutS in the mismatch recognition and directions for future investigation of mismatch repair.

• BMB Reports
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• v.35 no.2
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• pp.248-250
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• 2002

Discrimination between the amplification of mRNA and contaminating genomic DNA is a common problem when performing a reverse transcriptase-polymerase chain reaction (RT-PCR). Even after treatment of the samples with DNAse, it is possible that negative controls (samples in which no reverse transcriptase was added) will give positive results. This indicates that there was amplification of DNA, which was not generated during the reverse transcriptase step. The possibility exists that Taq DNA polymerase acts as a reverse transcriptase, generating cDNA from RNA during the PCR step. In order to test this hypothesis, we incubated samples with a DNAse-free RNAse after the cDNA synthesis. Comparison of the results that were obtained from these samples (incubated with or without DNAse-free RNAse) confirms that the reverse transcriptase activity of Taq DNA polymerase I is a possible source of false positive results when performing RT-PCR from intronless genes. Moreover, we describe here a simple and rapid method to overcome the false positive results that originate by this activity of Taq polymerase.

• Proceedings of the Korean Vacuum Society Conference
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• 2013.08a
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• pp.92.1-92.1
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• 2013

Molecular electronics has been the subject of intese research for many years because of the fundamental interest in molecular charge transport and potential applications, such as (bio)nanosensors and molecular memory devices. Molecular electronics requires a method for making reliable eletrical contacts to singlemolecules. To date, several approaches have been reported: scanning-probe microscopy, mechanical break junctions, nano patterning, and direct deposition of electrode on a self-assembled monolayers. However, most methods are laborious and difficult for large-scale application and more importantly, cannot control the number of moleucles in the junction. Recently, DNA has been used as a template for metallic nanostructures (e.g., Ag, Pd, and Au nanowires) through DNA metallization process. Furthermore, oligodeoxynucleotides have been tethered to organic molecules by using conventional organic reactions. Collectively, these techniques should provide an efficient route toward reliable and reproducible molecular electronic devices with large-scale fabrication. Therefore, I will present a paradigm for the fabrication of moleuclar electronic devices by using micrometer-sized DNA-singe organic molecule and DNA triblock structures.

• Archives of Pharmacal Research
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• v.20 no.3
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• pp.212-217
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• 1997

Induction of an adaptive response to ionizing radiation in mouse lymphoma (EL4) cells was studied by using cell survival fraction and apoptotic nucleosomal DNA fragmentation as biological end points. Cells in early log phase were pre-exposed to low dose of ${\gamma}$-rays (0.01 Gy) 4 or 20 hrs prior to high dose ${\gamma}$-ray (4, 8 and 12 Gy for cell survival fraction analysis; 8 Gy for DNA fragmentation analysis) irradiation. Then cell survival fractions and the extent of DNA fragmentation were measured. Significant adaptive response, increase in cell survival fraction and decrease in the extent of DNA fragmentation were induced when low and high dose .gamma.-ray irradiation time interval was 4 hr. Addition of protein or RNA synthesis inhibitor, cycloheximide or 5,6-dichloro-1-.betha.-d-ribofuranosylbenzimidazole (DRFB), respectively during adaptation period, the period from low dose ${\gamma}$-ray irradiation to high dose ${\gamma}$-ray irradiation, was able to inhibit the induction of adaptive response, which is the reduction of the extent DNA fragmentation in irradiated EL4 cells. These data suggest that the induction of adaptive response to ionizing radiation in EL4 cells required both protein and RNA synthesis.

• Korean Journal of Microbiology
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• v.7 no.1
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• pp.10-21
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• 1969

The effect of glucose and 2-thiobarbituric acid on the biosynthesis of cell constituents such as protein, carbohydrate, DNA, RNA, phospholipid and PCA-soluble phosphate compounds in Chlorella duing the life cycle was measured, and the changes in the content of these main cellular components of the algal cell were analyzed in connection with the nuclear and cytoplasmic divison. In the normal autotrophic synchronous culture the contents of protein, RNA, and DNA in the cell showed a chracteristic changes according to the progress of cell development, increasing more or less throughout all the life cycle. The synthesis of protein is more prominent in the division period nad that of DNA is more active in the ripening period, while the synthesis of RNA is more rapid in the growing and ripening periods than other developmental stages. The period of division cycle was little affected by glucose in the medium, although the synchrony of the growth and cellular division was disturbed and the n value increased. The cotents of protein, carbohydrate, RNA nad DNA of the cell were increased by the glucose treatment throughout all the life cycle. On the other hand, both of cellular growth and division were retarded severely and the n value was decreased by the 2-thiobarbituric acid treatment throughout all the life cycle. On the other hand, both of cellular growth and division were retarded severely and the n value was decreased by the 2-thiobarbituric acid treatment. The synthesis of protein, carbohydrate, DNA, RNA and phospholipid of the cell was also retarded by 2-thiobarbituric acid. In the autotrophic, mixotrophic and 2-thiobarbituric acid-treated cultures, each having different mode cytoplasmic division, a common general schema occurring in the cell during the life cycle may be drawn as follows. The ratio of RNA to protein attains maximum value in the $L_1$-cell stage prior to the nuclear division and thereafter decreases during the periods of ripening and division. The ratio of PCA-soluble phosphate compounds to protein increased from the begining of the culture to $L_4$-cell stage successively and thereafter decreased gradually during the division period, while the ratio of protein to DNA kept almost constant up to the division period and thereafter increased during the division period. Therefore, it is presumed that the increase in the ratio of RNA to protein is to be an inducer of nuclear division and that the cytoplasmic division is induced by the increase in the ratio of protein to DNA.

• BMB Reports
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• v.40 no.2
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• pp.156-162
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• 2007

Two lysis-defective but DNA synthesis non-defective temperature-sensitive (ts) mutants of mycobacteriophage L1, L1G23ts23 and L1G25ts889 were found to be defective also in phage-specific RNA synthesis in the late period of their growth at 42$^{\circ}C$each to the extent of 50% of that at 32$^{\circ}C$The double mutant, L1G23ts23G25ts889 showed the ts defect in phage RNA synthesis that was nearly additive of those shown individually by the two single-mutant parents. Both G23 and G25 were shown to start functioning sometimes between 30 and 45 min after infection but the former gene might be dispensable after 45 min, while the latter was not. Northern analysis also shows that at 42$^{\circ}C$>, L1G23ts23 affects RNA synthesis more strongly than L1G25ts889 from L1 DNA segments that serve as the template for late gene transcription. Among the 21 virion and 12 non-virion late proteins synthesized by L1, L1G23ts23 is defective in the synthesis of at least 9 virion and all of non-virion proteins at 42$^{\circ}C$>. In contrast, L1G25ts889 is completely defective in synthesis of all the 33 late proteins. Possible roles of G23 and G25 in the positive regulation of transcription of different sets of late genes of L1 have been discussed.