• Title/Summary/Keyword: linker-scanning

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Linker Scanning Analysis of the BPV-1 Upstream Regulatory Region

  • Kim, Hee-Dai;Rho, Jae-Rang;Choe, Joon-Ho
    • BMB Reports
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    • v.28 no.4
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    • pp.368-373
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    • 1995
  • The upstream regulatory region (URR) of bovine papillomavirus type 1 (BPV) contains promoters and a conditional transcriptional enhancer that is trans-activated by the viral E2 protein. After deleting the 5' and 3' ends of BPV URR, BamHI linkers were inserted into several positions of BPV URR without causing an addition or a deletion of URR sequences. Most linker scanning mutations did not show any effects on the transcription of P7940 and P89 promoters in BPV URR. However, several mutants showed reduced transcriptional activities. Based on our results we found that the AP-2 and Sp1 binding sites were important for basal level transcription of BPV URR in the absence of the E2 protein and that the CTF/NF-1 site is dispensable for E2 transactivation of BPV URR transcription.

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Focal adhesion linker proteins expression of fibroblast related to adhesion in response to different transmucosal abutment surfaces

  • Moon, Yeon-Hee;Yoon, Mi-Kyeong;Moon, Jung-Sun;Kang, Jee-Hae;Kim, Sun-Hun;Yang, Hong-Seo;Kim, Min-Seok
    • The Journal of Advanced Prosthodontics
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    • v.5 no.3
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    • pp.341-350
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    • 2013
  • PURPOSE. To evaluate adherence of human gingival fibroblasts (HGFs) to transmucosal abutment of dental implant with different surface conditions with time and to investigate the roles of focal adhesion linker proteins (FALPs) involved in HGFs adhesion to abutment surfaces. MATERIALS AND METHODS. Morphologies of cultured HGFs on titanium and ceramic discs with different surface were observed by scanning electron microscopy. Biocompatibility and focal adhesion were evaluated by ultrasonic wave application and cell viability assay. FALPs expression levels were assessed by RT-PCR and western blot. RESULTS. There seemed to be little difference in biocompatibility and adhesion strength of HGFs depending on the surface conditions and materials. In all experimental groups, the number of cells remaining on the disc surface after ultrasonic wave application increased more than 2 times at 3 days after seeding compared to 1-day cultured cells and this continued until 7 days of culture. FALPs expression levels, especially of vinculin and paxillin, also increased in 5-day cultured cells compared to 1-day cultured fibroblasts on the disc surface. CONCLUSION. These results might suggest that the strength of adhesion of fibroblasts to transmucosal abutment surfaces increases with time and it seemed to be related to expressions of FALPs.

Molecular Linker Enhanced Assembly of CdSe/ZnS Core-Shell Quantum Dots (분자 끈을 활용한 CdSe/ZnS 양자 점의 향상된 배열)

  • Cho, Geun Tae;Lee, Jong Hyeon;Nam, Hye Jin;Jung, Duk Young
    • Korean Chemical Engineering Research
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    • v.46 no.6
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    • pp.1081-1086
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    • 2008
  • QDs-LEDs(quantum dot light emitting device) should contain well-organized arrays of QDs on an electron transport layer. Thin films of CdSe/ZnS core-shell QDs were successfully fabricated on $TiO_2$ substrates by using PDMS stamp and micro contact printing method. 2-Carboxyethylphosphonic acid(CAPO) and 1,6-hexanedithiol(HDT) were employed as molecular linkers in assembling CdSe/ZnS core-shell QDs with high-density and uniform array. The CAPO increased the binding strength between the QDs and the substrates, and the HDT induced the strong inter-particle attractions of assembled QDs. The assembling properties of QDs thin films were characterized by SEM, AFM, optical microscope and photoluminescence spectroscope(PL).

Analysis of Porcine $\beta$-casein Gene Promoter by Site-directed Mutagenesis

  • Chung, Hee-Kyoung;Seong, Hwan-Hoo;Im, Seok-Ki;Lee, Hyun-Gi;Kim, Soon-Jeung;Lee, Poongyeong;Lee, Yun-Keun;Chang, Won-Kyong;Moosik Kwon
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.71-71
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    • 2002
  • Promoters for milk proteins have been used far producing transgenic animals due to their temporal and spatial expression patterns. ${\beta}$-casein, a calcium-sensitive casein, is a major milk protein that corresponds ca. 30 per cent of total milk protein. Expression of ${\beta}$-casein is controlled by lactogenic hormones such as prolactin (PRL), composite response elements (CoREs) and transcription factors. CoREs are clusters of transcription factor binding sites containing both positive and negative regulatory elements. ${\beta}$-casein gene promoter contains various regions (CoREs) for gene transcription. We analyzed the promoter region by mutagenesis using exonuclease III and linker-scanning. Transcription control elements usually are positioned in 5'-flanking region of the gene. However, in some cases, these elements are located in other regions such as intron 1. The nucleotide sequences of ${\beta}$-casein promote. region has been reported (E12614). However, the properties of the promoter is not yet clear. In this study, we plan to investigate the properties of cis-regulating elements of porcine ${\beta}$-casein by mutation analysis and expression analysis using dual-luciferase repoter assay system.

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Preparation of Highly Cross-linked, Monodisperse Poly(methyl methacrylate) Microspheres by Dispersion Polymerization; Part I. Batch Processes

  • Lee, Ki-Chang;Lee, Sang-Yun
    • Macromolecular Research
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    • v.15 no.3
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    • pp.244-255
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    • 2007
  • Nucleation is the most sensitive stage in the preparation of highly cross-linked, monodisperse microspheres by dispersion polymerization, since the addition of a small amount of cross-linker results in particle deformation and coagulation. To overcome these problems, $5\;{\mu}m$ poly(methyl methacrylate) seed particles prepared by dispersion polymerization were used in the preparation of mono disperse, cross-linked PMMA particles containing up to 7 wt% divinylbenzene by seeded batch dispersion polymerization. Spherical particles with a narrow size distribution containing up to 8 wt% of EGDMA were prepared by seeded multi-batch dispersion polymerization processes. These particles were identified by scanning electron microscopy and DSC.

Synthesis of Montmorillonite/Poly(acrylic acid-co-2-acrylamido-2-methyl-1-propane sulfonic acid) Superabsorbent Composite and the Study of its Adsorption

  • Zhu, Linhui;Zhang, Lili;Tang, Yaoji
    • Bulletin of the Korean Chemical Society
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    • v.33 no.5
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    • pp.1669-1674
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    • 2012
  • A novel superabsorbent composite was prepared by intercalation polymerization of acrylic acid (AA) and 2-acrylamido-2-methyl-1-propane sulfonic acid (AMPS) in the presence of montmorillonite (MMT), using ammonium persulfate (APS) as an initiator and $N,N'$-methylenebisacrylamide (MBA) as a cross linker. The superabsorbent composite was characterized by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), X-ray diffraction (XRD) and thermogravimetric analysis (TGA). Maximum absorbency of the composite in distilled water and 0.9% sodium chloride solution was 722 and 108 g/g, respectively. The composite was used for removal of heavy metal ions from aqueous solutions. Maximum amount of adsorption for $Ni^{2+}$, $Cu^{2+}$ and $Pb^{2+}$ was 211.0, 159.6 and 1646.0 mg/g, respectively, and the adsorption was in accordance with both Langmuir and Freundlich model. The composite could be regenerated and reused in wastewater treatment.

Poly(Dimethylaminoethyl Methacrylate)-Based pH-Responsive Hydrogels Regulate Doxorubicin Release at Acidic Condition

  • Lee, Seung-Hun;You, Jin-Oh
    • Journal of the Korean Applied Science and Technology
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    • v.32 no.2
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    • pp.202-214
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    • 2015
  • Stimuli-responsive biomaterials that alter their function through sensing local molecular cues may enable technological advances in the fields of drug delivery, gene delivery, actuators, biosensors, and tissue engineering. In this research, pH-responsive hydrogel which is comprised of dimethylaminoethyl methacylate (DMAEMA) and 2-hydroxyethyl methacrylate (HEMA) was synthesized for the effective delivery of doxorubicin (Dox) to breast cancer cells. Cancer and tumor tissues show a lower extracellular pH than normal tissues. DMAEMA/HEMA hydrogels showed significant sensitivity by small pH changes and each formulation of hydrogels was examined by scanning electron microscopy, mechanical test, equilibrium mass swelling, controlled Dox release, and cytotoxicity. High swelling ratios and Dox release were obtained at low pH buffer condition, low cross-linker concentration, and high content of DMAEMA. Dox release was accelerated to 67.3% at pH 5.5 for 6-h incubation at $37^{\circ}C$, while it was limited to 13.8% at pH7.4 at the same time and temperature. Cell toxicity results to breast cancer cells indicate that pH-responsive DMAEMA/HEMA hydrogels may be used as an efficient matrix for anti-cancer drug delivery with various transporting manners. Also, pH-responsive DMAEMA/HEMA hydrogels may be useful in therapeutic treatment which is required a triggered release at low pH range such as gene delivery, ischemia, and diabetic ketoacidosis.

Applications of Transposon-Based Gene Delivery System in Bacteria

  • Choi, Kyoung-Hee;Kim, Kang-Ju
    • Journal of Microbiology and Biotechnology
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    • v.19 no.3
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    • pp.217-228
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    • 2009
  • Mobile genetic segments, or transposons, are also referred to as jumping genes as they can shift from one position in the genome to another, thus inducing a chromosomal mutation. According to the target site-specificity of the transposon during a transposition event, the result is either the insertion of a gene of interest at a specific chromosomal site, or the creation of knockout mutants. The former situation includes the integration of conjugative transposons via site-specific recombination, several transposons preferring a target site of a conserved AT-rich sequence, and Tn7 being site-specifically inserted at attTn7, the downstream of the essential glmS gene. The latter situation is exploited for random mutagenesis in many prokaryotes, including IS (insertion sequence) elements, mariner, Mu, Tn3 derivatives (Tn4430 and Tn917), Tn5, modified Tn7, Tn10, Tn552, and Ty1, enabling a variety of genetic manipulations. Randomly inserted transposons have been previously employed for a variety of applications such as genetic footprinting, gene transcriptional and translational fusion, signature-tagged mutagenesis (STM), DNA or cDNA sequencing, transposon site hybridization (TraSH), and scanning linker mutagenesis (SLM). Therefore, transposon-mediated genetic engineering is a valuable discipline for the study of bacterial physiology and pathogenesis in living hosts.

High capacity polymer for nickel determination in environmental samples

  • Panahi, Homayon Ahmad;Feizbakhsh, Alireza;Dadjoo, Fatemeh;Moniri, Elham
    • Advances in environmental research
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    • v.2 no.4
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    • pp.309-321
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    • 2013
  • High AA new high capacity sorbent for preconcentration and determination of nickel in environmental samples was synthesized. The sorbent was synthesized by copolymerization of allyl glaycidyl ether / imminodiacetic acid with N,N-dimethylacrylamide as functional monomers in the presence of N,N-bismethylenacryl amid as cross linker and characterized by Fourier transform infra red spectroscopy, elemental analysis, thermogravimetric analysis and scanning electron microscopy. A recovery of 93.6% was obtained for the metal ion with 0.1 M, sulfuric acid as the eluting agent. The sorption capacity of the functionalized sorbent was 55.9 $mgg^{-1}$. The equilibrium sorption data of Ni(II) on polymeric sorbent were analyzed using Langmuir, Freundlich, Temkin and Redlich.Peterson models. Based on equilibrium adsorption data the Langmuir, Freundlich and Temkin constants were determined 0.87 (L mg-1), 25.87 ($mgg^{-1}$) $(Lmg^{-1})^{1/n}$ and 171.4 ($Jmol^{-1}$) respectively at pH 4.5 and $20^{\circ}C$.

Chitosan Nanoparticles as a New Delivery System for the Anti-HIV Drug Zidovudine

  • Dahmane, El Montassir;Rhazi, Mohammed;Taourirte, Moha
    • Bulletin of the Korean Chemical Society
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    • v.34 no.5
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    • pp.1333-1338
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    • 2013
  • Chitosan-based nanoparticles (CSNP) were prepared through ionic cross-linking and gelation of chitosan (CS) by tripolyphosphate (TPP). CS properties such as molecular weight, and preparation conditions were screened and the resulting nanoparticles were examined by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The obtained particles were consistently spherical with an overall diameter of approximately $107{\pm}20$ nm. They were successfully used as a carrier for Zidovudine, an anti-human immunodeficiency virus (HIV) which, to our knowledge, is novel. The encapsulation ability, loading capacity, and controlled release behavior for these CSNP was evaluated. Results indicated that their intrinsic properties were strongly affected by properties inherent to CS such as molecular weight, and by the preparation condition, such as cross-linking density, which depends on the concentration of the cross-linker. In vitro release tests for the entrapped zidovudine showed that the CNNP provided a continuous release that can last upwards 20 h.