• Title/Summary/Keyword: chimeric

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Inhibitory Effects of Chimeric Decoy Oligodeoxynucleotide in the Regulation of Transcription Factors NF-κB and Sp1 in an Animal Model of Liver Cirrhosis (간경화 동물모델에서 Chimeric decoy oligodeoxynucleotide로 억제되는 NF-κB와 Sp1 전사인자 발현 억제 효과에 대한 연구)

  • Kim, Kyung-Hyun;Park, Ji-Hyun;Kim, Soo-Jung;Lee, Woo-Ram;Chang, Young-Chae;Kim, Hyun-Chul;Park, Kwan-Kyu
    • Journal of Life Science
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    • v.19 no.10
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    • pp.1360-1367
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    • 2009
  • Liver fibrosis is a process of healing and scarring in response to chronic liver injury. Following injury, an acute inflammation response takes place resulting in moderate cell necrosis and extracellular matrix damage. To develop a novel therapeutic approach in hepatic fibrogenesis, we examined the simultaneous suppression of the transcription factors NF-$\kappa$B and Sp1, which regulate acute inflammation and continuous deposition of extracellular matrix in liver fibrosis. We employed chimeric decoy oligodeoxynucleotide containing the consensus sequences of both NF-$\kappa$B and Sp1 binding sites, to suppress these transcription factors simultaneously. Treatment of chimeric decoy oligodeoxynucleotide reduced the activity of hepatic stellate cells in vitro, and decreased the expression of fibrotic and proinflammatory gene responses in a mouse model of liver fibrosis. These results suggest that chimeric decoy oligodeoxynucleotide strategy can be a potential therapeutic application to prevent liver fibrosis.

Effect of escherichia coli plasmid DNA sequences on plasmid replication in yeast (효모에서 plasmid의 복제에 대장균 plasmid DNA가 미치는 영향에 관한 연구)

  • 김태국;최철용;노현모
    • Korean Journal of Microbiology
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    • v.27 no.1
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    • pp.16-20
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    • 1989
  • The effect of E. coli plasmid DNA sequences contained by chimeric vectors on plasmid replication was investigated. We constructed YRp7- or 2.$\mu$m circle-based plasmids containing E. coli plasmid DNA sequences and those not containing it. By examining their maintenance in yeast, we showed that plasmid without E. coli plasmid DNA sdquences was nore stable and presented higher copy number, and espressed higher level of hepatitis B viral surface antigen as a foreign gene. This result suggested that E. coli plasmid DNA sequences within chimeric plasmid somehow inhibited plasmid replication in yeast.

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Production of Transgenic Chimeric Chickens Using Blastodermal Cells

  • Yan, Haifeng;Lee, Chaeyoung;Xiao, Bingnan;Trefil, Pavel;Liu, Shixun;Kim, Younyoung;Wu, Xiaolin
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.2
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    • pp.158-164
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    • 2005
  • A practical approach was proposed to produce transgenic chimeric chickens using blastodermal cells (BCs). The chicken BCs were mechanically dissociated and transferred into the recipient eggs that had been exposed to 500 rads irradiation of$^{60}Co$ and windowed on the equatorial plane. Chimeric chickens were generated using two models: the crosses (MXL) from Black Minors (ii,EE,b/b) ♂${\times}$Barred Leghorns (ii,ee,B/-) ♀ as donors and White Leghorns (WL, II) as acceptors (Model 1), or the Black Heifengs (BH, ii,EE,bb) as donors and Hua-xing white (HW, II) as recipients (Model 2). The treated eggs were incubated in their original shells in normal conditions until hatching. Green fluorescent protein (GFP) gene was transferred into the BCs derived from MXL and BH via lipofectamine and the pEGFP-C1, and transfection efficiency into the BCs was examined under a fluorescent microscope. Potential transgenic chimeras were selected based on the proposed methods in this study. Using the fresh BCs, the best rate of phenotypic chimeras was 6.7% and 26.0% in model-1 groups, and model-2 groups, respectively. We also described the optimized conditions for transfection. Although 30% of the BCs transfected in vitro emitted green light under an inverted fluorescent microscope, no embryos injected with the transfected BCs expressed foreign GFP gene at 3-4 days.

Bi-functional Activities of Chimeric Lysozymes Constructed by Domain Swapping between Bacteriophage T7 and K11 Lysozymes

  • Alcantara, Ethel H.;Kim, Dong-Hee;Do, Su-Il;Lee, Sang-Soo
    • BMB Reports
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    • v.40 no.4
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    • pp.539-546
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    • 2007
  • The lysozymes encoded by bacteriophage T7 and K11 are both bifunctional enzymes sharing an extensive sequence homology (75%). The constructions of chimeric lysozymes were carried out by swapping the N-terminal and C-terminal domains between phage T7 and K11 lysozymes. This technique generated two chimeras, T7K11-lysozyme (N-terminal T7 domain and C-terminal K11 domain) and K11T7-lysozyme (N-terminal K11 domain and C-terminal T7 domain), which are both enzymatically active. The amidase activity of T7K11-lysozyme is comparable with the parental enzymes while K11T7-lysozyme exhibits an activity that is approximately 45% greater than the wild-type lysozymes. Moreover, these chimeric constructs have optimum pH of 7.2-7.4 similar to the parental lysozymes but exhibit greater thermal stabilities. On the other hand, the chimeras inhibit transcription comparable with the parental lysozymes depending on the source of their N-terminals. Taken together, our results indicated that domain swapping technique localizes the N-terminal region as the domain responsible for the transcription inhibition specificity of the wild type T7 and K11 lysozymes. Furthermore, we were able to develop a simple and rapid purification scheme in purifying both the wild-type and chimeric lysozymes.

Expression of Cinnamic Acid 4-Hydroxylase Chimeric Gene fused with Sesquiterpene Cyclase Promoter from Hot Pepper in Tobacco (고추의 sesquiterpene cyclase promoter-cinnamic acid 4-hydroxylase chimeric gene의 담배에서 발현)

  • 이경민;윤용휘;김길웅;이인중;신동현
    • Journal of Life Science
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    • v.14 no.4
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    • pp.657-663
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    • 2004
  • Tobacco transformants harboring cinnamic acid 4-hydroxylase gene (C4H) fused with susquiterpene cyclase promoter was developed in order to regulate biosynthesis of phenolic compounds by the expression of the introduced gene. Twenty transformants for each specific promoter were used to analyze the incorporation of the chimeric genes by PCR and Southern blot analysis. PCR products of NPTII(neomycin phosphotransferase) gene (553bp) were detected in the transgenic tobacco plants. The incorporation of the chimeric gene was confirmed in the Southern blot analysis. C4H activity in the transgenic plants was elevated by UV-irradiation and its level was higher compared to that of control plants.

Immunization of mice with chimeric protein-loaded aluminum hydroxide and selenium nanoparticles induces reduction of Brucella melitensis infection in mice

  • Tahereh Goudarzi;Morteza Abkar;Zahra Zamanzadeh;Mahdi Fasihi-Ramandi
    • Clinical and Experimental Vaccine Research
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    • v.12 no.4
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    • pp.304-312
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    • 2023
  • Purpose: Due to the many problems with commercially available vaccines, the production of effective vaccines against brucellosis is a necessity. The aim of this study was to evaluate the immune responses caused by the chimeric protein consisting of trigger factor, Bp26, and Omp31 (TBO) along with aluminum hydroxide (AH/TBO) and selenium (Se/TBO) nanoparticles (NPs) as adjuvants in mouse model. Materials and Methods: Recombinant antigen expression was induced in Escherichia coli BL21 (DE3) bacteria using IPTG (isopropyl-d-1-thiogalactopyranoside). Purification and characterization of recombinant protein was conducted through NiFe3O4 NPs, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and Western blot. NP characteristics, including morphology and particle size, were measured in vitro. The recombinant TBO was loaded on to AH and Se NPs and were administered subcutaneously. After mice immunization, measurement of antibody titter and protection assay was performed. Results: The average sizes of AH and Se NPs were about 60 nm and 150 nm, respectively. The enzyme-linked immunosorbent assay results showed that the serum of mice immunized by subcutaneous injection with both nanovaccines produced significant immunoglobulin G (IgG) responses against the chimeric antigen. The results of TBO-specific IgG isotype (IgG2a/IgG1) analysis showed that both AH and Se NPs induced a type to T-helper immune response. In addition, the results of the challenge with the pathogenic strain of Brucella melitensis 16M showed that vaccinated mice with AH/TBO NPs indicated a higher reduction of bacterial culture than immunized mice with Se/TBO NPs and TBO alone. Conclusion: The results showed that AH NPs carrying chimeric antigen can be a promising vaccine candidate against brucellosis by producing protective immunity.

Ig G fusion 단백질을 사용한 리간드-수용체의 상호작용

  • 천혜경
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.11a
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    • pp.143-145
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    • 1994
  • Chimeric fusion proteins involving IgG have proven valuable in studying protein-protein interactions and may possess therapeutic applications as well. For example, three receptor subtypes for the natriuretic peptides, when fused to the Fc portion of human IgG ${\gamma}$ chain, were quantitatively and qualitatively indistinguishable from the native receptor, thus allowing detailed structure-function studies of the receptor. In an attempt to block human immunodeficiency virus infectivity with soluble derivatives of CD4, a CD4/IgG Fc chimeric molecule was shown to increase the plasma half life of soluble CD4 and possessed the added advantage of IgG Fc-mediated placental transfer. In the case of the KGFR, this approach provided a framework for dissection of its ligand binding domains and made it possible to demonstrate that high affinity binding sites for two ligands, aFGF and KGF, reside within different receptor Ig-like domains. Chimeric molecules fused to immunoglobulins would have the advantages of secretion from transfected cells as well as detection and purification from medium utilizing Staphylococcus aureus Protein A. In addition, where highly related receptors make their discrimination very hard due to the difficulties in generating specific immunochemical probes, IgG fusion protein with tailor-made specificities confers particular advantages to elucidate patterns of receptor distribution and expression. The approach described here may have general applications in defining ligand-receptor interactions as well as searching for specific agonists and antagonists of receptor function.

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Defective Interfering HIV-1 Pseudotypes Carrying Chimeric CD4 Protein

  • Park, Seung-Won;Ye, Zhiping;Schubert, Manfred;Paik, Soon-Young
    • BMB Reports
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    • v.34 no.6
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    • pp.566-572
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    • 2001
  • Chimeric CD4 proteins were assembled. They contained the entire CD4 ectodomain that is linked to different membrane anchors. Membrane anchors consisted of either glucosyl phosphatidyl inositol (gpi), the transmembrane and cytoplasmic regions of HIV-1 Env protein, or the vesicular stomatitis virus G glycoprotein, respectively. The HIV-1 co-receptor CXCR4 and CD4 were independently inserted into viral envelopes. We compared the insertion of six different CD4/CXCR4 constructs into HIV-1 envelopes, as well as their functionality in targeting and specific infection of cells that constitutively express the HIV-1 Env protein. All of the six different HIV-1 (CD4/CXCR4) pseudotypes were able to transduce Env (+) cells at similar efficiency. In addition, stable transduction of the Env (+) recipient cells demonstrated that all chimeric proteins were functional as receptors for Env when inserted into HIV-1 envelopes. In fact, these results demonstrate for the first time a stable transduction by a targeted HIV-1 pseudotype virus.

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