• Title/Summary/Keyword: DNA Synthesis

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A Productive Replication of Hyphantria cunea Nucleopolyhedrovirus in Lymantria dispar Cell Line

  • Demir, Ismail;Demirbag, Zihni
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1485-1490
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    • 2006
  • In this study, comparative replicational properties of Hyphantria cunea nucleopolyhedrovirus (HycuNPV) in Lymantria dispar (IPLB-LdElta) and Spodoptera frugiperda (IPLB-Sf21) cell lines were investigated. Our microscopic observations showed that cytopathic effects (CPEs) in LdElta cells appeared 12 h later than those in Sf21 cells. Whereas polyhedral inclusion bodies (PIBs) formed at 48 h postinfection (p.i.) in LdElta cells, it formed at 36 h p.i. in Sf21 cells. Extracellular virus production determined according to the 50% tissue culture infective dose ($TCID_{50}$) method in LdElta cells started about 12 h later when compared with Sf21 cells. Titers of extracellular virus in LdElta and Sf21 cells were calculated as $1.77{\times}10^9$ plaque forming units (PFU)/ml and $5.6{\times}10^9PFU/ml$, respectively, at 72 h p.i. We also showed that viral DNA replication began at 12 h p.i. in both cell lines. Viral protein synthesis was determined by SDS-polyacrylamide gel electrophoresis (PAGE) and polyhedrin synthesis was observed at 12 h p.i. in both cell lines. The results indicate that while the synthesis of macromolecules is 12 h later and production of extracellular virus is almost 3-fold lower in LdElta cells compared with those in Sf21 cells, the LdElta cell line is still a productive cell line for infection of HycuNPV.

Effects of FIS Protein on rnpB Transcription in Escherichia coli

  • Choi, Hyun-Sook;Kim, Kwang-sun;Park, Jeong Won;Jung, Young Hwan;Lee, Younghoon
    • Molecules and Cells
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    • v.19 no.2
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    • pp.239-245
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    • 2005
  • Factor for inversion stimulation (FIS), the Escherichia coli protein, is a positive regulator of the transcription of genes that encode stable RNA species, such as rRNA and tRNA. Transcription of the rnpB gene encoding M1 RNA, the catalytic subunit of E. coli RNase P, rapidly declines under stringent conditions, as does that of other stable RNAs. There are multiple putative FIS binding sites upstream of the rnpB promoter. We tested whether FIS binds to these sites, and if so, how it affects rnpB transcription. In vitro binding assays revealed specific binding of FIS to multiple sites in the rnpB promoter region. Interestingly, FIS bound not only to the upstream region of the promoter, but also to the region from +4 to +18. FIS activated rnpB transcription in vitro, but the level of activation was much lower than that of the rrnB promoter for rRNA. We also examined the effects of FIS on rnpB transcription in vivo using isogenic $fis^+$ and $fis^-$ strains. rnpB transcription was higher in the $fis^-$ than the $fis^+$ cells during the transitions from lag to exponential phase, and from exponential to stationary phase.

Effects of the Protein Fraction of Panax ginseng on Primary Cultured Chicken Brain Cells and DRG (인삼 단백분획물이 일차배양한 계배의 뇌세포 및 DRG에 미치는 영향)

  • Park, Mi-Jung;Song, Jin-Ho;Kim, Sun-Yeou;Kim, Young-Choong
    • YAKHAK HOEJI
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    • v.34 no.5
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    • pp.365-373
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    • 1990
  • The effects of the protein fraction of Panax ginseng on primary cultured chicken embryonic brain cells and DRG cultured with a deficient medium were studied. The protein fraction was further fractionated into four groups according to the molecular weight; larger than 10,000 dalton(fraction A), between 5,000 and 10,000 daltons(fraction B), between 1,000 and 5,000 daltons(fraction C), between 500 and 1,000 daltons(fraction D). All four protein fractions at the concentration of $100\;{\mu}g/ml$ significantly increased the number of the brain cells which promoted the neurite outgrowth. The activity of PDHC in the brain cells was elevated significantly by the protein fraction B at the concentration of $100\;{\mu}g/ml$. It was noted that $100\;{\mu}g/ml$ protein fraction C and D significantly enhanced the synthesis of protein in the brain cells. At the concentration of $100\;{\mu}g/ml$, the protein fraction B enhanced RNA synthesis and the protein fraction A significantly enhanced DNA synthesis in the brain cells. The protein fractions B, C, and D significantly promoted the neurite outgrowth of DRG at the concentration of $100\;{\mu}g/ml$.

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Polyamine Prevent Apoptotic Cell Death by Regulation of Apoptosis Related Gene Expression in Porcine Parthenotes

  • Cui, Xiang-Shun;Jin, Yong-Xun;Hwang, Kyu-Chan;Kim, Nam-Hyung
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.230-230
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    • 2004
  • Polyamines, namely putrescine, spermidine, and spermine, are biogenic low-molecular-weight aliphatic amines. Polyamines play important roles in DNA stabilization, RNA and protein synthesis, membrane stabilization, modulation of ion channels, and protection against oxygen radicals and are essential for cell homeostasis, cell growth, and tumorigenesis. (omitted)

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Mutational analsysis of phage SP6 transcription initiation and a new transcription vector

  • Kang, Changwong;Nam, Sang-Chul;Lee, In-Woo
    • The Microorganisms and Industry
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    • v.14 no.1
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    • pp.7-11
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    • 1988
  • Efficient in vitro RNA synthesis can be easily accomplished from cloned DNA using bactrio-phage SP6, T7 or T3 RNA polymerase. Despite its popularity as in vitro transcription system, molecular mechanisms of bacteriophage transcription has not been studied, although physical and catalytic properties of several phage RNA polymerases have well been documented (1). Only recently the T7 promoter has been physically mapped by footprinting of the T7 RNA polymerase (2,3). These simple phage systems, however, could be useful for detailed molecular studies of transcription.

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Synthesis and characterization of transferrin-polyethylenimine conjugate for targeted gene delivery

  • Lee, Kyung-Man;Kim, In-Sook;Shin, Sang-Chul;Oh, In-Joon
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.315.2-316
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    • 2003
  • Polyethylenimine (PEI) has been used as a non-viral gene delivery carrier. To improve the efficacy of transfection, transferrin was incorporated by covalent linkage to PEI. As a model plasmid DNA, pHME185/b-gal, a mammalian expression vector was used. The transferrin-polyethylenimine (TfPEI) was synthesized by conjugate PEI with transferrin using sodium periodateand and characterized by FT-IR and 1H-NMR. (omitted)

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