generation of fluorescent igg fusion proteins in mammalian cells
Báo cáo khoa học: Functional characterization of ecdysone receptor gene switches in mammalian cells docx
... DNA binding domain and VP16 activation domain Various combinations of the EcR ligand binding domain, VP16 activation domain and GAL4 DNA binding domain were constructed (Fig 1A) and tested in NIH ... probably in uence the behavior of fusion protein upon binding to ligand Apparently, having activation domain (VP16) and DNA binding domain (GAL4), in that order, on the NH2-terminal end of EcR ... EcR fusion proteins require heterodimeric partner for binding to ligand as well as for transactivation of reporter genes in cells An EcR gene switch that includes a fusion protein containing...
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Expression studies of the proteolytic p10 fragment of the neuronal cdk5 activator in mammalian cells
... Neuro-2a and p10 fusion proteins were overexpressed in these cells The influences of p10 overexpression in these mammalian cells were subsequently monitored and studied using a number of biotechniques ... accumulation of aging pigments in nondividing cells, the decrease of striatal dopamine receptors, and the proliferation of smooth muscle cells in blood vessel walls and so on The identification of the ... which is defined by cell swelling and bursting, releasing intracellular contents which may damage surrounding cells and often cause inflammation, apoptotic cells not release constituents into extracellular...
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Expression studies of the proteolytic p10 fragment of the neuronal cdk5 activator in mammalian cells
... Neuro-2a and p10 fusion proteins were overexpressed in these cells The influences of p10 overexpression in these mammalian cells were subsequently monitored and studied using a number of biotechniques ... accumulation of aging pigments in nondividing cells, the decrease of striatal dopamine receptors, and the proliferation of smooth muscle cells in blood vessel walls and so on The identification of the ... which is defined by cell swelling and bursting, releasing intracellular contents which may damage surrounding cells and often cause inflammation, apoptotic cells not release constituents into extracellular...
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Intein mediated generation of n terminal cysteine proteins and their applications in live cell bioimaging and protein microarray
... labeling of N-terminal Cys proteins for Bioimaging 30 3.2.1 In vitro labeling 30 3.2.2 Expression and in vivo cleavage of intein -fusion to generate 33 N-terminal Cys proteins in bacteria and mammalian ... N-terminal Cys containing proteins at the C-terminus of the Ssp DnaB mini intein (17kDa) using the pTWIN vector (NEB, USA) Intein-mediated cleavage occurs between the last amino acid of the mini intein ... were inserted in frame after the C-terminal of the Ssp DnaB intein (Intein1) in the pTWIN1 and pTWIN2 vectors, with the first amino acid of the protein of interest engineered to be a cysteine...
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Tài liệu Báo cáo khoa học: DNA modification with cisplatin affects sequence-specific DNA binding of p53 and p73 proteins in a target site-dependent manner pptx
... interaction with either of the p73 isoforms; at rb ¼ 0.06, only slight (10–15%) inhibition of binding was detected Thus, the p73d and p73b proteins exhibited behavior upon binding to cisplatin-treated ... p53–DNA interaction (or even prevention of target recognition by the protein) DNA binding of the C-terminally truncated p53(1–363) protein Fig The effects of DNA modification with cisplatin on binding ... protein was decreased due to formation of cisplatin adducts within the p53 DNA-binding site (p53DBS), resulting in increased p53 binding to the labeled probe The pPGM4 fragment (c) contains cisplatinresistant...
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Tài liệu Báo cáo khoa học: Extended half-life upon binding of destabilized intrabodies allows specific detection of antigen in mammalian cells pdf
... maintained in the cells by binding to antigen Generation of soluble binding-defective and cysteine-deleted mutants To demonstrate that the observed intracellular colocalization was the result of ... 7A-cotransfected cells, thus demonstrating that strong in vivo binding of the destabilized 13R4-d1-EGFP proteins allows them to be maintained in the cells To rule out the possibility that the EGFP part of ... measuring protein mobility and activity in living cells Nature Cell Biol 3, 145–147 Bachmair A, Finley D & Varshavsky AK (1986) In vivo half-life of a protein is a function of its amino-terminal...
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Tài liệu Báo cáo khoa học: Enhanced expression of Mcm proteins in cancer cells derived from uterine cervix docx
... by hematoxylin and eosin staining We examined the expression of these proteins in a cancer-free layer of squamous epithelial cells, carcinoma in situ (CIS) and invasive cancer, all of which were ... significance of Mcm proteins in the aberrant proliferation of cancer cells In addition, Mcm3 and proteins seem to be useful as a marker to discriminate cancer cells and dysplastic cells in the uterine ... more abundant in HeLa cells than in WI-38 cells Synthesis of Mcm4, and proteins was accelerated in HeLa cells compared to WI-38 cells, but the turnover rate of the Mcm4 protein within 22 h did...
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Báo cáo khoa học: Site-specific phosphorylation of MCM4 during the cell cycle in mammalian cells pot
... eukaryotic cells including human cells [7] Targets of the kinase in the regulation of DNA replication include ORC2, Cdc6 and Mcm proteins in Saccharomyces cerevisiae [8], and disregulation of these proteins ... prevent binding of MCM complex to chromatin [12] In contrast, a recent finding suggests that an intermediate level of phosphorylation of MCM4 is required for chromatin binding during interphase ... the presence of geminin, which inhibits the ability of CDT1 to load MCM proteins onto chromatin, may help prevent re-replication of DNA in the G2 and M phase During interphase, chromatin-bound MCM4...
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Báo cáo khoa học: Identification of sodium salicylate as an hsp inducer using a simple screening system for stress response modulators in mammalian cells pptx
... examined the effects of SA on stress response in mammalian cells using a simple screening system, and revealed that SA is a potent Hsp inducer in mammalian cells, thereby protecting cells against ... transcription of hsp genes and the accumulation of Hsps in various mammalian cells and a concomitant increase of thermoresistance of cells Thus, SA may be used for the protection of cells against deleterious ... containing a geneticin resistant gene (3 lg) were cotransfected into C3H10T1/2 cells (1 · 107 cells per 100 mm dish) with 30 lL lipofectAMINE reagent (Invitrogen) according to the manufacturer’s instructions,...
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Báo cáo Y học: Mydj2 as a potent partner of hsc70 in mammalian cells doc
... R.A., Narain, Y., Rankin, J & Rubinsztein, D.C (2000) Effects of heat shock, heat shock protein 40 (HDJ-2) and proteosome inhibition on protein aggregation in cellular models of Huntington’s disease ... to bind to immobilized Mydj2-His protein (lanes 3, 3¢) In contrast, no binding between the immobilized dj2 protein and the inducible hsp70 protein was observed (Fig 6, lanes: 3, 3¢) Interestingly, ... for and analyzed by 10% SDS-mini PAGE and Western blotting Indirect immunofluorence CV1 cells growing in coverslips were incubated at 37 °C or 43 °C for h as indicated in the text and figure legends...
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Báo cáo khoa học: Differential expression of liver and kidney proteins in a mouse model for primary hyperoxaluria type I pdf
... 11 Neuhoff V, Arold N, Taube D & Ehrhardt W (1988) Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie ... enzyme supports the induction of fatty acid metabolism The reduction in d-amino acid oxidase expression in the kidney proteome is interesting, in view of the contribution of this enzyme to glyoxylate ... Phosphoglycerate mutase Indolethylamine N-methyltransferase Peroxiredoxin Apolipoprotein A-I Abhydrolase domain containing 14b D-Amino Protein name ApoA4 ActB Nsfl1C Acy1 Pgam1 Inmt Prdx6 ApoA1 Abhd14b...
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Báo cáo khoa học: Activity of the plant peptide aglycin in mammalian systems potx
... of a binding protein in membrane extracts tested P, pancreas; L, liver; K, kidney; M, muscle Upward arrows indicate beginning of injection of the extract, downward arrows indicate beginning of ... trypsin inhibitor, lgÆmL)1 aprotinin, lgÆmL)1 leupeptin and mgÆmL)1 bacitracin), disintegrated in a JJ-2 homogenizer (GuoHua Instrument Co., Wuhan, China) with two volumes of buffer A containing ... characterization of aglycin binding proteins Biosensor analysis Spreeta biosensor and software (American TI Corp., Attleboro, MA, USA) was used to detect aglycin binding proteins Aglycin was immobilized...
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Báo cáo Y học: Regulation of peptide-chain elongation in mammalian cells pptx
... calmodulin binding and the GxGxxG motif that may be involved in substrate binding (see text) within this motif prevents calmodulin binding and thus also activity [43] Removal of the C-terminus (residues ... sources Instead, activation of PKA results in a marked rise in the maximal activity of eEF2 kinase from ventricular myocytes, suggesting that these cells may contain a distinct isoform of eEF2 kinase, ... existence of multiple forms of eEF2 kinase in phaeochromocytoma cells and the regulatory properties of eEF2 kinase in ventricular cardiomyocytes suggest the existence of a distinct form in these cells...
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Báo cáo khoa học: Mitochondrial oxidative stress causes mitochondrial fragmentation via differential modulation of mitochondrial fission–fusion proteins potx
... activity in the cells overexpressing Drp1 was much lower than that in non-overexpressing cells, whereas the activity of caspase-9 in Mfn2-overexpressing cells was higher than that in non-overexpressing ... the involvement of Drp1 in HF-LPLI-induced apoptosis at a fluence of 120 JÆcm)2 in ASTC-a-1 cells Both control cells and HF-LPLI-treated cells were labeled with MitoTracker, and endogenous Drp1 in ... Geneva) for kindly providing pshRNA Drp1, R J Youle (National Institutes of Health) for kindly providing pYFP–Mfn2 and pYFP–Drp1, and M Miura (Riken Brain Science Institute) for kindly providing pSCAT9...
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Báo cáo khoa học: Expression studies of the core+1 protein of the hepatitis C virus 1a in mammalian cells pot
... core+1 protein in mammalian cells have failed, including transfection of cells with plasmid DNA encoding the core sequence or infection with recombinant herpes simplex virus expressing the core–E1–E2 ... that its detection in mammalian cells and in the HCV infectious system is elusive This study shows that intracellular levels of the core+1 protein in mammalian cells are strongly in uenced not only ... S–myc-expressing plasmid (pHPI-1496) and increasing amounts of the core-expressing vector (pHPI-1499) (Fig 2A), in the Expression of the HCV-1 core+1 protein presence of MG-132 Immunoblotting indicated...
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Báo cáo hóa học: "Photochemically reduced polyoxometalate assisted generation of silver and gold nanoparticles in composite films: a single step route" pptx
... spectra of reduced composite at various time intervals The intensity of 750 band increased with an increase in the time of irradiation indicating that more silicotunsgstic acid is getting reduced ... the intensity of the band at 481 nm increased which might be due to the increase in the amount of Ag (Fig 9b) When increasing the concentration of the metal nanoparticles in composite film the intensity ... dipping time of 30 are shown in Fig The particle size as well as the amount of Au in the composite is found to increase with the increase in the concentration of HAuCl4 solution The change in...
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Báo cáo hóa học: " A Generalized Algorithm for the Generation of Correlated Rayleigh Fading Envelopes in Wireless Channels" ppt
... limits the rate of changes of fading in time domain, and consequently, results in the autocorrelation properties of fading Therefore, the algorithm generating Rayleigh fading envelopes in realistic ... Rayleigh fading envelopes in multipath, flat fading channels in a single time instant We can repeat steps and of this algorithm to generate Rayleigh envelopes in the continuous time interval It ... model of a Rayleigh fading generator for generating an individual baseband Rayleigh fading envelope proposed in [10, 16] is shown in Figure This model generates a Rayleigh fading envelope using inverse...
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Báo cáo y học: "Expression of bioactive bone morphogenetic proteins in the subacromial bursa of patients with chronic degeneration of the rotator cuff" pptx
... (green) and BMP-7 images of bone morphogenetic proteins Indirect immunofluorescence (red) in two areas of inflamed bursa (BMP)-2 (green) and BMP-7 (red) in two areas of inflamed bursa (a) and (b) ... Safranin O staining, an indicator of large deposits of proteoglycan, was negative (not shown) This is in agreement with the absence of mRNA for aggrecan in this tissue (see below) Within these ... tendon tissue (d) In all cases, treatment of the sections with 0.5 mM levamisole eradicated the enzyme reaction, leaving behind only staining in granules within individual cells that appeared...
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Báo cáo y học: "Identification of secondary targets of N-containing bisphosphonates in mammalian cells via parallel competition analysis of the barcoded yeast deletion collection" ppsx
... a haploinsufficient profile in yeast in the presence of N-BPs Among these was ALF1, a homolog of the mammalian tubulin cofactor B (TBCB) gene, which encodes the α-tubulin folding protein It has ... the initiation of the DNA replication by activating the minichromosome maintenance (MCM) protein Both DBF4-Cdc7 and MCM proteins are phosphorylated by the protein kinases ATM and ATR [25] Cells ... Rogers MJ: Nitrogen-containing bisphosphonates inhibit the mevalonate pathway and prevent post-translational prenylation of GTP-binding proteins, including Ras J Bone Miner Res 1998, 13:581-589...
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Báo cáo y học: "Autoprocessing of human immunodeficiency virus type 1 protease miniprecursor fusions in mammalian cells" pptx
... the GST-miniprecursor fusions are competent for autoprocessing in mammalian cells Darunavir and Indinavir inhibit fusion precursor autoprocessing in transfected 293T cells To further examine autoprocessing ... proximal site) In the present study, a similar fusion construct was engineered for mammalian expression to examine protease autoprocessing in transfected mammalian cells Autoprocessing of the fusion ... antibodies In the presence of cell-permeable Darunavir and Indinavir [26,27], precursor autoprocessing was inhibited in a dose-dependent manner, as indicated by the appearance of increasing amounts of...
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