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Wyszukujesz frazę "fusion protein" wg kryterium: Temat

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    Tytuł:
    A systematic investigation of the stability of green fluorescent protein fusion proteins
    Autorzy:
    Janczak, Monika
    Bukowski, Michał
    Górecki, Andrzej
    Dubin, Grzegorz
    Dubin, Adam
    Wladyka, Benedykt
    Powiązania:
    https://bibliotekanauki.pl/articles/1038973.pdf
    Data publikacji:
    2015
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    crystallography
    fusion protein
    recombinant protein
    toxin-antitoxin system
    Opis:
    X-ray crystallography provides important insights into structure-function relationship in biomolecules. However, protein crystals are usually hard to obtain which hinders our understanding of multiple important processes. Crystallization requires large amount of protein sample, whereas recombinant proteins are often unstable or insoluble. Green fluorescent protein (GFP) fusion is one of the approaches to increase protein synthesis, solubility and stability, facilitating crystallization. In this study we analyze the influence of the linker length, composition and the position of GFP relative to the fusion partner on the fusion protein production and stability. To this end, multiple constructs of enzymatically impaired variant of PemKSa toxin from Staphylococcus aureus CH91 fused to GFP were generated. Fusion protein production in Escherichia coli was evaluated. The proteins were purified and their stability tested. PemKSa-α14aa-GFP fusion provided best production and stability. Obtained results demonstrate the importance of optimization of fusion protein construct, including linker selection and the order of fusion partners, in obtaining high quantities of stable protein for crystallization.
    Źródło:
    Acta Biochimica Polonica; 2015, 62, 3; 407-411
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    A new strategy: high level expression and immunogenicity analysis of triplicate repeated multigenes in Mycoplasma hyopneumoniae
    Autorzy:
    Li, J.
    Wang, G.
    Powiązania:
    https://bibliotekanauki.pl/articles/16647587.pdf
    Data publikacji:
    2023
    Wydawca:
    Polska Akademia Nauk. Czasopisma i Monografie PAN
    Tematy:
    multi-epitope fusion protein
    Mycoplasma hyopneumoniae
    new generation vaccines
    Opis:
    Highly immunogenic nucleotide fragments from 3 genes of Mycoplasma hyopneumoniae strain 232 were selected using information software technology. After repeating each fragment three times, a total of 9 nucleotide fragments were joined together to form a new nucleotide sequence called Mhp2321092bp. Mhp2321092bp was directly synthesized and cloned into a pET100 vector and expressed in Escherichia coli. After purification, the proteins were successfully validated by SDS-PAGE and Western blotting using mouse His-tag antibody and pig anti-Mhp serum. BALB/c mice were intraperitoneally injected with purified proteins in the high-dose (100 µg), medium-dose group (50 µg) and low-dose (10 µg) groups. Mice in each group were injected on day 1, day 8 and day 15 of feeding, respectively. Serum samples were collected from all mice on the day before immunization and on day 22 after immunization. The antibody level in the mouse serum was detected using western blotting using purified expressed proteins as antigens. IL-2, TNF-α and IFN-γ were simultaneously detected in the mouse serum by ELISA. The results showed that the 60 kDa protein was successfully expressed and reacted specifically with the specific serum Mhp His-Tag mouse monoclonal antibody and pig anti-Mhp serum. From day 0 to day 22 of immunization, IFN-γ increased from 269.52 to 467.74 pg/mL, IL-2 increased from 14.03 to 145.16 pg/mL, and TNF-α increased from 6.86 to 12.37 pg/mL. The IgG antibody in mice increased significantly from 0 day to day 22 after immunization. This study suggests that the expressed recombinant protein may serve as one of the novel vaccine candidates for Mhp.
    Źródło:
    Polish Journal of Veterinary Sciences; 2023, 26, 2; 275-283
    1505-1773
    Pojawia się w:
    Polish Journal of Veterinary Sciences
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    The location of fusion protein AUX1-YFP in a transgenic callus of Arabidopsis thaliana
    Autorzy:
    Fautynowicz, L.
    Tretyn, A.
    Wisniewska, J.
    Powiązania:
    https://bibliotekanauki.pl/articles/951189.pdf
    Data publikacji:
    2015
    Wydawca:
    Polska Akademia Nauk. Czytelnia Czasopism PAN
    Tematy:
    fusion protein
    auxin
    Arabidopsis thaliana
    P-glycoprotein
    indole-3-acetic acid
    Źródło:
    BioTechnologia. Journal of Biotechnology Computational Biology and Bionanotechnology; 2015, 96, 1
    0860-7796
    Pojawia się w:
    BioTechnologia. Journal of Biotechnology Computational Biology and Bionanotechnology
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    Expression of bovine leukemia virus protein p24 in Escherichia coli and its use in the immunoblotting assay.
    Autorzy:
    Bicka, Leokadia
    Kuźmak, Jacek
    Kozaczyńska, Bożena
    Płucienniczak, Andrzej
    Skorupska, Anna
    Powiązania:
    https://bibliotekanauki.pl/articles/1044191.pdf
    Data publikacji:
    2001
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    bovine leukemia virus
    fusion protein p24
    immunoblotting assay
    gag gene cloning
    Opis:
    The gag gene encoded protein, p24 of bovine leukemia virus (BLV), was cloned and expressed as thioredoxin-6xHis-p24 protein in Escherichia coli. The bacterial cells carrying plasmid pT7THis-p24 expressed the protein of 38 kDa that was detected by immunoblotting analysis using anti-p24 monoclonal antibodies and sera from BLV infected cattle and sheep. The purified p24 fusion protein was shown to be sensitive and specific for detection of BLV antibodies in the infected cattle.
    Źródło:
    Acta Biochimica Polonica; 2001, 48, 1; 227-232
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    Interleukin-6 biology is coordinated by membrane bound and soluble receptors.
    Autorzy:
    Rose-John, Stefan
    Powiązania:
    https://bibliotekanauki.pl/articles/1043433.pdf
    Data publikacji:
    2003
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    inflammation
    cytokine
    sgp130Fc fusion protein
    gp130
    cytokine receptor
    soluble receptor
    Opis:
    Cytokine receptors exist in membrane bound and soluble form. Both forms bind their ligands with comparable affinity. While most soluble receptors are antagonists in that they compete for the ligands with their membrane counterparts, some soluble receptors are agonists. In this case, the complex of ligand and soluble receptor binds on target cells to a second receptor subunit and initiates signal transduction. Soluble receptors of the IL-6 family of cytokines are agonists. In vivo, the IL-6/soluble IL-6R complex stimulates several types of target cells not stimulated by IL-6 alone, since they do not express the membrane bound IL-6R. This process has been named transsignaling. We have shown that in several chronic inflammatory diseases like chronic inflammatory bowl disease, peritonitis and rheumatoid arthritis, transsignaling via the soluble IL-6R complexed to IL-6 is a crucial point in the transition from the acute to the chronic state of the disease. The mechanism by which the IL-6/ soluble IL-6R complex regulates the inflammatory state is discussed.
    Źródło:
    Acta Biochimica Polonica; 2003, 50, 3; 603-611
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    Biochemical and structural studies of plant circadian clock proteins
    Autorzy:
    Saini, R.
    Szpotkowski, K.
    Kozak, M.
    Jaskolski, M.
    Davis, S.J.
    Powiązania:
    https://bibliotekanauki.pl/articles/80840.pdf
    Data publikacji:
    2013
    Wydawca:
    Polska Akademia Nauk. Czytelnia Czasopism PAN
    Tematy:
    circadian clock
    diurnal change
    plant circadian clock
    protein
    multiple feedback loop
    crystallization
    recombinant fusion protein
    early flowering
    small angle X-ray scattering
    protein-protein interaction
    biochemical study
    structural study
    Źródło:
    BioTechnologia. Journal of Biotechnology Computational Biology and Bionanotechnology; 2013, 94, 1
    0860-7796
    Pojawia się w:
    BioTechnologia. Journal of Biotechnology Computational Biology and Bionanotechnology
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    Immunogenicity of recombinant bacterial antigens expressed as fusion proteins in transgenic rice seeds
    Autorzy:
    Zaman, S.
    Islam, S.M.T.
    Khan, M.K.
    Alam, M.M.
    Uddin, M.I.
    Baby, N.I.
    Islam, S.
    Bhuiyan, T.R.
    Qadri, F.
    Seraj, Z.I.
    Powiązania:
    https://bibliotekanauki.pl/articles/80383.pdf
    Data publikacji:
    2017
    Wydawca:
    Polska Akademia Nauk. Czytelnia Czasopism PAN
    Tematy:
    rice seed
    transgenic rice plant
    gene sequence
    fusion protein
    tuberculosis
    vaccine
    oral vaccine
    Ag85B antigen
    cholera
    bacterial antigen
    immunogenicity
    Źródło:
    BioTechnologia. Journal of Biotechnology Computational Biology and Bionanotechnology; 2017, 98, 4
    0860-7796
    Pojawia się w:
    BioTechnologia. Journal of Biotechnology Computational Biology and Bionanotechnology
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    Engineered resistance against proteinases
    Autorzy:
    Milner, Malgorzata
    Chroboczek, Jadwiga
    Zagorski-Ostoja, Wlodzimierz
    Powiązania:
    https://bibliotekanauki.pl/articles/1040935.pdf
    Data publikacji:
    2007
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    fusion proteins
    proteinase inhibitors
    protein protection
    Opis:
    Exogenous proteinase inhibitors are valuable and economically interesting protective biotechnological tools. We examined whether small proteinase inhibitors when fused to a selected target protein can protect the target from proteolytic degradation without simultaneously affecting the function and activity of the target domain. Two proteinase inhibitors were studied: a Kazal-type silk proteinase inhibitor (SPI2) from Galleria mellonella, and the Cucurbita maxima trypsin inhibitor I (CMTI I). Both inhibitors target serine proteinases, are small proteins with a compact structure stabilized by a network of disulfide bridges, and are expressed as free polypeptides in their natural surroundings. Four constructs were prepared: the gene for either of the inhibitors was ligated to the 5' end of the DNA encoding one or the other of two selected target proteins, the coat protein (CP) of Potato potyvirus Y or the Escherichia coli β-glucuronidase (GUS). CMTI I fused to the target proteins strongly hampered their functions. Moreover, the inhibitory activity of CMTI I was retained only when it was fused to the CP. In contrast, when fused to SPI2, specific features and functions of both target proteins were retained and the inhibitory activity of SPI2 was fully preserved. Measuring proteolysis in the presence or absence of either inhibitor, we demonstrated that proteinase inhibitors can protect target proteins used either free or as a fusion domain. Interestingly, their inhibitory efficiency was superior to that of a commercial inhibitor of serine proteinases, AEBSF.
    Źródło:
    Acta Biochimica Polonica; 2007, 54, 3; 523-536
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    MutS as a tool for mutation detection
    Autorzy:
    Stanisławska-Sachadyn, Anna
    Sachadyn, Paweł
    Powiązania:
    https://bibliotekanauki.pl/articles/1041359.pdf
    Data publikacji:
    2005
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    fusion
    SNP
    chimeric protein
    MutS
    mutation detection
    Opis:
    MutS, a DNA mismatch-binding protein, seems to be a promising tool for mutation detection. We present three MutS based approaches to the detection of point mutations: DNA retardation, protection of mismatched DNA against exonuclease digestion, and chimeric MutS proteins. DNA retardation in polyacrylamide gels stained with SYBR-Gold allows mutation detection using 1-3 µg of Thermus thermophilus his6-MutS protein and 50-200 ng of a PCR product. The method enables the search for a broad range of mutations: from single up to several nucleotide, as mutations over three nucleotides could be detected in electrophoresis without MutS, due to the mobility shift caused by large insertion/deletion loops in heteroduplex DNA. The binding of DNA mismatches by MutS protects the complexed DNA against exonuclease digestion. The direct addition of the fluorescent dye, SYBR-Gold, allows mutation detection in a single-tube assay. The limited efficiency of T4 DNA polymerase as an exonuclease hampers the application of the method in practice. The assay required 300-400 ng of PCR products in the range of 200-700 bp and 1-3 µg of MutS. MutS binding to mismatched DNA immobilised on a solid phase can be observed thanks to the activity of a reporter domain linked to MutS. We obtained chimeric bifunctional proteins consisting of T. thermophilus MutS and reporter domains, like β-galactosidase or GFP. Very low detection limits for β-galactosidase could theoretically enable mutation detection not only by the examination of PCR products, but even of genomic DNA.
    Źródło:
    Acta Biochimica Polonica; 2005, 52, 3; 575-583
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    Coronaviruses fusion with the membrane and entry to the host cell
    Autorzy:
    Wędrowska, E.
    Wandtke, T.
    Senderek, T.
    Piskorska, E.
    Kopiński, P.
    Powiązania:
    https://bibliotekanauki.pl/articles/2085559.pdf
    Data publikacji:
    2020
    Wydawca:
    Instytut Medycyny Wsi
    Tematy:
    coronavirus
    spike protein
    membrane fusion
    viral entry
    nonstructural proteins
    replication complex
    Opis:
    Introduction. Coronaviruses (CoVs) are positive-strand RNA viruses with the largest genome among all RNA viruses. They are able to infect many host, such as mammals or birds. Whereas CoVs were identified 1930s, they became known again in 2003 as the agents of the Severe Acute Respiratory Syndrome (SARS). The spike protein is thought to be essential in the process of CoVs entry, because it is associated with the binding to the receptor on the host cell. It is also involved in cell tropism and pathogenesis. Receptor recognition is the crucial step in the infection. CoVs are able to bind a variety of receptors, although the selection of receptor remains unclear. Coronaviruses were initially believed to enter cells by fusion with the plasma membrane. Further studies demonstrated that many of them involve endocytosis through clathrin-dependent, caveolae-dependent, clathrin-independent, as well as caveolae-independent mechanisms. Objectives. The aim of this review is to summarise current knowledge about coronaviruses, focussing especially on CoVs entry into the host cell. Advances in understanding coronaviruses replication strategy and the functioning of the replicative structures are also highlighted. The development of host-directed antiviral therapy seems to be a promising way to treat infections with SARS-CoV or other pathogenic coronaviruses. There is still much to be discovered in the inventory of pro-and anti-viral host factors relevant for CoVs replication. The latest pandemic danger, originating from China, has given our previously prepared work even more of topicality.
    Źródło:
    Annals of Agricultural and Environmental Medicine; 2020, 27, 2; 175-183
    1232-1966
    Pojawia się w:
    Annals of Agricultural and Environmental Medicine
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
      Wyświetlanie 1-10 z 10

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