preparation of cell free extract
Báo cáo khoa học: Functional analysis of cell-free-produced human endothelin B receptor reveals transmembrane segment 1 as an essential area for ET-1 binding and homodimer formation pptx
... bacterial, yeast or insect cells is still complicated and often inefficient [5–11] Continuous exchange cell- free (CECF) expression systems based on Escherichia coli cell extracts have recently been ... after conventional expression in insect cells Ligand binding of cell- free endothelin B receptor A Single-particle analysis of CF-produced ETBcHx The quality of CF-expressed and purified ETBcHx was ... affinity to purified ETB in Brij78 micelles, as Ligand binding of cell- free endothelin B receptor indicated by the determined KD of nm, which is even lower than the value of 29 nm previously determined...
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báo cáo khoa học: "PVP-coated silver nanoparticles block the transmission of cell-free and cell-associated HIV-1 in human cervical culture" ppsx
... cytotoxic in only 20% of the cells of the cervical explant, whereas a dose of 0.6 mg/mL was cytotoxic to 23% of the cells PVPcoated AgNPs formulated in Replens gel inhibited cell viability by 5%, ... 18% of cells (Fig 2b) Raft-media has many antibiotics, which, when combined, result in cytotoxicity Inhibition of cell- free and cell- associated HIV-1 viral infection in the presence or absence of ... percentages of cells with tissue showing signs of inhibition of HIV-1 infection transmission were evaluated with respect to the positive control The cytopathic effects of the viral infection of MT2 cells...
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Báo cáo y học: " Prognostic value of cell-free plasma DNA in patients with cardiac arrest outside the hospital: an observational cohort study" pdf
... Page of 11 sample to measure cell- free plasma DNA was taken from the antecubital vein of each patient immediately after return of spontaneous circulation in the emergency room Plasma and cells ... GE/ml with a sensitivity of 63%, specificity of 69%, positive likelihood ratio of 1.75 (95% CI, 1.44 to 2.35) and correct classification rate of 62% The best cutoff value of six-hour lactate in ... computed tomography ratio of 2.41 (95% CI, 2.04 to 3.26) and correct classification rate of 73% Regarding the secondary endpoint of inhospital mortality, the best cut-off value of plasma DNA was 3,485...
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Rheological aspect of cell free layer formation in micro blood flow experimental and numerical study
... Physiological implication of CFL 13 Overview of dissertation 15 CHAPTER II: A COMPARATIVE STUDY OF HISTOGRAM-BASED THRESHOLDING METHOD FOR DETERMINATION OF CELLFREE LAYER WIDTH IN ... study on the suspension of blood, which includes blood rheology, cell mechanics and physiology Blood, a vital fluid of humans, is a concentrated suspension of cells and cell fragments in plasma ... in the circulation The normal shape of RBC consists of a biconcave discoid with a width of ~7.7 μm, thickness of ~2.8 μm, surface area of ~130 μm2 and volume of ~98 μm3 [37] Due to its shape, RBC...
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Báo cáo khoa học: 15 N-Labelled proteins by cell-free protein synthesis Strategies for high-throughput NMR studies of proteins and protein–ligand complexes doc
... strains Cell- free protein synthesis systems are far more inert with regard to isotope scrambling because the pool of metabolic enzymes present in the cell extract is not regenerated Thus, cell extracts ... N-labelled proteins by cell- free synthesis K Ozawa et al Fig Combinatorial isotope labelling scheme Oval symbols identify the 15N-labelled amino acids used in the cell- free preparation of the five different ... [15N]-labelling of selected residues was suppressed by the addition of amino acids at natural isotopic abundance [17] In the case of cell- free protein synthesis, however, the costs of the [15N]labelled...
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Báo cáo khoa học: The influence of cold shock proteins on transcription and translation studied in cell-free model systems pdf
... fractions free of nucleic acids were concentrated by ultraltration to a concentration of 5.9 mm From L of cell culture, mg of BsCSP could be prepared The molar extinction coefcient of BsCSP was ... mRNA Therefore free mRNA accumulates in these cell- free systems [13] and excessive secondary structure formation takes place Results Effect of cold shock proteins on the combined cell- free transcription/translation ... western blot with a-Ras Cell- free expression of H-Ras encoded on pET14brasc in h batch reaction under the inuence of different concentrations of TmCSP as indicated Ten microlitres of each reaction...
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Báo cáo khoa học: Optimization of an Escherichia coli system for cell-free synthesis of selectively 15N-labelled proteins for rapid analysis by NMR spectroscopy pdf
... 2004 Cell- free synthesis of 15 N-labelled proteins (Eur J Biochem 271) 4089 4090 K Ozawa et al (Eur J Biochem 271) Fig (Continued) Ó FEBS 2004 Ó FEBS 2004 Cell- free synthesis of The cell- free ... Optimization of other conditions for in vitro protein synthesis Fig Cell- free synthesis of PpiB under control of phage k promoters Identical volumes of reaction products were loaded into lanes of a 15% ... concentrations of MgCl2 and template DNA were found to vary with different batches of S30 extract Concentration of the S30 extract by dialysis against a solution of PEG 8000 [2] reduced the volume of the...
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Báo cáo khoa học: Efficient synthesis of a disulfide-containing protein through a batch cell-free system from wheat germ pdf
... the crude cell- free protein synthesis solution The present biotinylation technique may be useful for preparation of biotin-labeled proteins and also for nonradioisotope detection of cell- free synthesized ... formation of disulfide bonds in E coli cell- free protein synthesis systems [15,16] The active fraction of the scFv produced in the E coli cell- free system with chaperones was nearly half of the synthesized ... antigen the extract was passed through a gel filtration column just before the start of reaction With the use of this low-DTT extract, the scFvLH mRNA was translated in the presence of [14C]Leu,...
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Báo cáo khoa học: The consensus motif for N-myristoylation of plant proteins in a wheat germ cell-free translation system ppt
... efficiency of protein N-myristoylation and the identity of the amino acids at positions 3, and of the consensus motif in plant target proteins with the use of a Cell- free N-myristoylation of plant ... digestion and extraction with the use of an XL-trypKit (Promega) The extracted peptides were desalt- Cell- free N-myristoylation of plant proteins ed and concentrated with the use of a ZipTip C18 ... previous finding of an inhibitory effect of Pro at position of full-length translation products on MAP function in the wheat germ cell- free translation system [35] We also detected low levels of [35S]Met...
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Báo cáo khóa học: High level cell-free expression and specific labeling of integral membrane proteins doc
... Ó FEBS 2004 Cell- free expression of membrane proteins (Eur J Biochem 271) 569 more than 10 h [6,7] Essential components of the cell- free system such as the bacterial S30 extract preparation, ... Aliquots of the cell- free extract were frozen in liquid nitrogen and stored at )80 °C The cell- free expression was performed in the continuous exchange mode using a membrane with a cutoff of 15 ... Klammt et al (Eur J Biochem 271) Fig Cell- free production of membrane proteins Lanes and 2, in vivo expression Samples of total cell extracts containing 10 lg of protein were analysed by SDS/PAGE...
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Báo cáo khóa học: Effects of Escherichia coli ribosomal protein S12 mutations on cell-free protein synthesis doc
... cultures of the wild-type and S12 mutants were inoculated into 2· YT medium (16 g of tryptone, 10 g of yeast extract, and g of NaCl per L) The concentration of streptomycin used for cultivation of ... Cell- free translation Preparation of rpsL mutants Spontaneous streptomycin-resistant or streptomycin-dependent mutants of E coli BL21 were obtained from colonies that grew within days after cells ... ratio of the incorporation of [3H]Leu to that of [14C]Phe Results Growth characteristics of S12 mutants We isolated 150 colonies of E coli BL21, grown on plates containing various concentrations of...
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Báo cáo sinh học: " Stimulation of poliovirus RNA synthesis and virus maturation in a HeLa cell-free in vitro translation-RNA replication system by viral protein 3CDpro" docx
... stocks with a 1:1 mixture of phenol and chloroform The purified RNA was precipitated by the addition of volumes of ethanol Preparation of HeLa cytoplamic extracts HeLa S10 extracts were prepared ... separation of 5S protomers and 14S pentamers The amount of radioactivity at the bottom of the tubes of the gradients was not determined (A) Comparison of samples obtained in the absence or presence of ... cytoplasm of the infected cell These complexes provide a suitable environment for increased local concentration of all the viral and cellular proteins needed for RNA replication and encapsidation of...
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Báo cáo y học: " Free radical scavenging activity and lipoxygenase inhibition of Mahonia aquifolium extract and isoquinoline alkaloids" ppt
... w/w) The remaining aqueous extract was crude Mahonia extract Fractionation of stem-bark extract Mahoniae A solution of 25 % NH4OH was added to the remaining acidified extract so as to adjust its ... certificate and according to the Law of the National Council of Slovak Republic on the protection of animals No 115/1995, Edict of Ministry of Agriculture on the breeding of society animals, wild animals ... local commercial sources and they were of analytical grade quality Preparation and analysis of Mahoniae stem-bark extract Plant material The stem-bark of cultivated Mahonia aquifolium was collected...
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báo cáo khoa học: " A simple and high-sensitivity method for analysis of ubiquitination and polyubiquitination based on wheat cell-free protein synthesis" potx
... layer consisted of 60 A260 units of the wheat germ extract (CellFree Sciences, Yokohama, Japan) and μg creatine kinase (Roche Diagnostics K K., Tokyo, Japan) in 25 μl of SUB-AMIX® (CellFree Sciences) ... from three independent experiments Analysis of the Wheat Cell- free Based Ubiquitination in the Presence of Proteasome Inhibitor It is known that some cell extracts, such as rabbit reticulocyte or ... Identification of activity and features of E3 ligases offers important information about the ubiquitin-dependent regulation system Our luminescent method based on the wheat cell- free system produced...
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Chlorine free extraction of cellulose from rice husk and whisker isolation
... of RH and RH free of extractives a peak at 1734 cm−1 can be seen which can also be assigned to C O of ketone but due to hemicelluloses Fig also shows an ATR-FTIR spectrum of extractive -free cellulose ... structure of each compound In this work FTIR was employed with the aim of verifying if lignin and hemicelluloses were removed from the extracted cellulose In this work FTIR spectra of RH, RH free of extractives, ... Extraction of cellulose and preparation of nanocellulose from sisal fibers Cellulose, 15, 149–159 Orts, W J., Shey, J., Imam, S H., Glenn, G M., Guttman, M E., & Revol, J F (2005) Application of cellulose...
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Effect of red blood cell aggregation on arteriolar cell free layer formation and its physiological functions
... dynamics of cell- free layer formation 62 (e) Frequency distribution of cell- free layer variations 63 4.1.4 Discussion 66 (a) Relation between mean and SD of cell- free ... Relation between mean and SD of cell- free layer widths 59 (c) Effect of aggregation and flow reduction on mean and SD of cell- free layer widths 60 (d) Effect of aggregation and flow ... 67 (b) Effect of aggregation and flow rate on mean and SD of cell- free layer widths 68 (c) Cell- free layer variations in terms of frequency distribution and magnitude of deviation...
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CHEMICAL AND BIOLOGICAL CHARACTERIZATION OF LIGUSTICUM WALLICHII EXTRACT IN 3t3 l1 AND HEP g2 CELL LINES
... phase cells (45.36, 50.53 and 54.84%, respectively) 41 IPE 72 h Cell Number Cell Number 48 h Cell Number Cell Number 24 h Cell Number Cell Number Control Figure 3.4 DNA cell cycle histograms of ... effect of phthalides extracted from chuangxiong on Hep-G2 cell line To study the effect of phthalides extracted from chuangxiong in 3T3-L1 cell model as related to diabetes 5.3 Implications of This ... HPLC 3.2 Preparation of Crude Phthalides Extracts from Chuangxiong One gram of chuangxiong powder was subjected to 50 mL of methanol extraction under reflux condition (70 ºC) for h The extract...
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One step, ultrasonication mobilized, solvent free extractionsynthesis of nanocurcumin from turmeric
... to improve extraction efficiency and reduce extraction time [35] The proposed benefits of UAE include: (a) overall, enhancement of extraction yield or rate, (b) enhancement of aqueous extraction ... confirmation of the successful extraction of curcumin RSC Advances Page of 25 View Article Online DOI: 10.1039/C5RA06002H h (WBS 5) of sonication and -100% sonication energy (Fig 1(c)) of probe sonication ... technique showed higher extraction efficiency of curcumin in RSC Advances Page 12 of 25 View Article Online DOI: 10.1039/C5RA06002H Mechanism of sonication based extraction of curcumin from turmeric...
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Báo cáo y học: " Preparation of RGD-modified Long Circulating Liposome Loading Matrine, and its in vitro Anti-cancer Effects"
... of RGD-M-LCL and measurement of its particle size RGD-M-LCL was freeze-dried by a cryoprotectant of sugar in a sugar-lipid quality ratio of 2.0 (22), then was redissolved with DMEM The size of ... of the cancer cells (P
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