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Báo cáo y học: "HIV-1 Tat protein enhances Microtubule polymerization" docx
... 1) These values are higher than the turbidity value of 0.30 obtained with 15 µM tubulin alone (Table 1) Thus Tat from different HIV subtypes are able to enhance tubulin polymerization However, ... values) A high proportion of apoptotic cells (31 %) was also obtained with 10 µM of Tat Eli Furthermore, the percentage of hypodiploid cells obtained with 10 µM Tat Oyi is low (10.4 %) and is the same ... presence of paclitaxel (line 3) However, full depolymerization in the presence of paclitaxel was obtained with a longer incubation at 4°C (data not shown) The reversibility of following an incubation...
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Tài liệu Báo cáo khoa học: Tissue expression and biochemical characterization of human 2-amino 3-carboxymuconate 6-semialdehyde decarboxylase, a key enzyme in tryptophan catabolism pptx
... (here named ACMSD II) ACMSD II coding sequence was obtained by using the primers pair 4fw ⁄ 11 rev (Fig 2A) Again, two distinct PCR products were obtained, the longer (887 bp) corresponding to part ... the threshold cycle values obtained with 10-fold serial dilutions of linearized pGEM T easy vectors harboring the ACMSD variants Standard curves for 18S RNA were obtained using cDNAs from each ... respectively In both cases, proteins were expressed as inclusion bodies (not shown) Any effort to obtain soluble recombinant proteins, including lower growth temperatures (18 °C) and isopropyl b-D-1-thiogalactopyranoside...
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Tài liệu Báo cáo khoa học: Structural and biochemical characterization of a human adenovirus 2/12 penton base chimera pptx
... of the N-terminal region [16], and crystals of the hAd2 and hAd2 ⁄ 12 chimera dodecahedra were obtained with protein N-terminally truncated by 49 residues [19] The structure shows that the truncated ... (Carlsbad, CA, USA) Protease inhibitors were from Roche (Basel, Switzerland) and Ni-NTA resin was obtained from Bio-Rad (Hercules, CA, USA) The minifiber construct was a gift from the Laboratory of ... value was taken as a data point Titrations were continued until a stable anisotropy value was obtained Fluorescence anisotropy, A, is defined as A ¼ ðIV À IH Þ=ðIV þ 2IH Þ where IV and IH refer...
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Tài liệu Báo cáo khoa học: Molecular and biochemical characterization ofD-phosphoglycerate dehydrogenase fromEntamoeba histolytica A unique enteric protozoan parasite that possesses both phosphorylated and nonphosphorylated serine metabolic pathways docx
... family In addition, the GDH sequence was also available from E histolyitca [30] The alignment obtained was corrected by manual inspection, and unambiguously aligned 182 sites were selected and ... amino acid substitution process, assuming homogeneous rates across sites Based on the best tree obtained, a G-shape parameter (a) of the discrete G-distribution with eight categories that approximates ... tergcongensis, 31% to Bacillus anthracis, Bacillus cereus and Caulobacter crescentus, 27% to Bacillus subtilis and Escherichia coli, 24–26% to human, mouse, rat, Schizosaccharomyces pombe and Saccharomyces...
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Tài liệu Báo cáo Y học: Structural and biochemical characterization of neuronal calretinin domain I– II (residues 1– 100) Comparison to homologous calbindin D28k domain I–II (residues 1 –93) pdf
... 4749) The AUTOASSIGN program [37] was used to obtain automatic assignments By running the program with several sets of parameters, we were able to obtain a consistent auto-assignment for residues ... experiments were processed using FELIX (MSI, San Diego) The AUTOASSIGN program [37] was used to obtain automatic assignments All assignments were manually checked for residue type and compared ... pH 8.1 The stimulated-echo with longitudinal eddy current delay (STE-LED) method was applied to obtain diffusion ordered spectroscopy (DOSY) spectra at 278 K [43,44] We implemented Bruker’s LEDGS2S...
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Báo cáo Y học: Purification and biochemical characterization of some of the properties of recombinant human kynureninase pptx
... were harvested and active enzyme extracted A purification protocol was then developed (Table 1) to obtain homogeneous enzyme Purification protocol All steps were performed at °C and long-term storage ... 90 Ó FEBS 2002 Recombinant human kynureninase (Eur J Biochem 269) 2071 tryptic mass fingerprint obtained by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) MS of a band of ... CRICKETGRAPH and GraphPad PRISM3 software packages, and the kinetic parameters Km and Vmax were obtained using non-linear regression Lineweaver–Burk and Dixon [14] plots were used to characterize...
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Báo cáo Y học: Structural and biochemical characterization of calhepatin, an S100-like calcium-binding protein from the liver of lungfish (Lepidosiren paradoxa) docx
... Cu2+-binding-induced fluorescence change was corrected for nonspecific quenching by subtracting the values of a linear term obtained from the final portion of the Cu2+-binding curve, corresponding to fluorescence ... equation containing an additional linear term, but the value obtained for the association constant was indistinguishable from that obtained with corrected data RESULTS Purification of lungfish calhepatin ... concentration) fractions obtained from the Superose column were incubated for 12 h and applied again to the gel-filtration column In both cases, the two forms were obtained again and the ratio...
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Báo cáo khóa học: Identification of a gene encoding Lon protease from Brevibacillus thermoruber WR-249 and biochemical characterization of its thermostable recombinant enzyme pptx
... 35 Spacer ) 16 ) 10 Inverted repeat Ref Br thermoruber Br brevis B subtilis B subtilis B subtilis B subtilis B subtilis B subtilis E coli lon lon lon clpX ftsH spoIIG amyR rA consensus r70 consensus ... transcription of Bt- lon is enhanced after a shift to higher temperatures (data not shown) However, the mechanisms of induction of Bt- lon require more study Characterization of Bt- Lon To characterize Bt- Lon, ... Lon gene (Bt- lon) Based on the codon usage preference of thermophilic Br thermoruber WR-249, the following two degenerate Preparation of Bt- lon expression constructs The full-length Bt- lon flanked...
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Báo cáo khoa học: An a-proteobacterial type malate dehydrogenase may complement LDH function in Plasmodium falciparum Cloning and biochemical characterization of the enzyme potx
... and Pf MQO were also cloned to obtain the DNA probes for analysis of its expression in P falciparum cultures Pf LDH was also overexpressed in E coli cultures to obtain the enzyme for evaluation ... aligned sequences were selected manually and realigned to obtain proper alignment of all sequences PHYLIP format tree output was selected to obtain the bootstrap neighbor-joining tree An unrooted ... The crude model so obtained was then refined by minimization using an analogous approach reported previously [34] The structure of the Pf MDH in complex with OAA and NADH was obtained as follows...
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Báo cáo khoa học: Identification and biochemical characterization of the Anopheles gambiae 3-hydroxykynurenine transaminase pot
... mosquito-borne diseases by vector control Recombinant Ag-HKT bacterial expression and enzyme purification To obtain large amounts of highly purified A gambiae 3-HKT for subsequent enzymatic studies, we expressed ... specificity towards different amino acceptors Data are expressed as percentage of the activity obtained using glyoxylate as the a-ketoacid amino acceptor in the Ag-HKT catalysed transamination ... 330 nm and quantified by interpolating the experimental data in the corresponding standard curve obtained as above The kinetic parameters of Ag-HKT 5660 F Rossi et al catalysis towards d,l-3HK,...
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Báo cáo Y học: Agmatine oxidation by copper amine oxidase Biosynthesis and biochemical characterization of N-amidino-2-hydroxypyrrolidine pdf
... phenylmethanesulfonyl fluoride, and Escherichia coli lipopolysaccharide (serotype 0127:B8) were obtained from Sigma Chemical Co [3H]L-arginine (specific activity 2.0 TBqÆmmol)1) and [3H]clonidine ... NENTM Life Science Products (Boston, MA, USA) Deuterium oxide (99.8% isotopic enrichment) was obtained from Cortec (Paris, France) All the other chemicals were from Merck AG (Darmstadt, Germany) ... inhibitor (K ) for the P sativum copper m amine oxidase catalyzed oxidation of agmatine, were obtained from the dependence of the initial velocity for agmatine oxidation (vi) on the substrate...
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Báo cáo khóa học: Isolation and biochemical characterization of two soluble iron(III) reductases from Paracoccus denitrificans docx
... patterns similar to those found above were obtained (Fig 5B) This behavior indicates a sequential mechanism with NADH and FMN as substrates The results obtained on studying the Fe(III) reductase ... [27] Materials and methods Organism and cultivation Paracoccus denitrificans, strain CCM 982 was obtained in freeze-dried form from the Czech Collection of Microorganisms (CCM), Masaryk University, ... UV/visible spectrophotometer (Pharmacia) using a 1-cm light path cell Fluorescence spectra were obtained using an LS 50B spectrofluorimeter (Perkin Elmer) N-Terminal sequence analysis The highly...
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Identification and biochemical characterization of tetrahydrolipstatin targets in m bovis BCG at different metabolic states
... up-regulated during mycobacteria regrowth from non-replicating persistent (NRP) bacilli Therefore, to obtain a functional insight into mycobacterial esterases and/or lipases in mycobacterial NRP, we ... SDS Sodium dodecyl sulphate SEM Standard error of the mean TAG Triacylglycerol TB Tuberculosis TBTA Tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]amine TCEP Tris(2-carboxyethyl)phosphine TDM Trehalose ... reach the lungs, they are phagocytosed by alveolar macrophages The macrophages then invade the subtending epithelial layer, Madhu Sudhan Ravindran 27 which induces a localized inflammatory response...
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Biochemical characterization of thermophilic lignocellulose degrading enzymes and their potential for biomass bioprocessing
... characterization of cellulose- and xylan-degrading enzymes from a thermophilic microbial consortium obtained by enrichment of yard waste compost as a source Materials and methods 2.1 Chemicals and ... (Rockford, IL, USA), respectively 2.2 Lignocellulosic substrates Pine wood saw dust (PWSD) was obtained from a local saw mill in Rapid City, SD Corn stover (CS) and prairie cord grass (PCG) were ... (IJEE), Volume 2, Issue 1, 2011, pp.99-112 2.10 Ethanol fermentation Lignocellulosic hydrolyzates obtained following incubation of TMC on PCS and PPCG for 120 h served as feedstock for ethanol production...
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Tài liệu Báo cáo khoa học: Pyrimidine-specific ribonucleoside hydrolase from the archaeon Sulfolobus solfataricus – biochemical characterization and homology modeling doc
... NucleoSpin Plasmid kit for plasmid DNA preparation were obtained from Genenco (Duren, Germany) Specically synthesized oligodeoxyribonucleotides were obtained from MWG-Biotech (Ebersberg, Germany) Restriction ... purchased from Stratagene (La Jolla, CA, USA) Thermolysin was obtained from Boehringer (Mannheim, Germany) Trypsin, proteinase K, subtilisin, nucleosides, purine and pyrimidine bases, O-bromoacetyl-N-hydroxysuccinimide ... in molecular mass studies were obtained from Sigma (St Louis, MO, USA) IPTG was from Applichem (Darmstadt, Germany) Sephacryl S-200 and AH-Sepharose 4B were obtained from Amersham Pharmacia Biotech...
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Tài liệu Báo cáo khoa học: Biochemical characterization and inhibitor discovery of shikimate dehydrogenase from Helicobacter pylori docx
... spectropolarimeter with a 1-mm path-length cuvette at 25 °C Experimental data were corrected by subtracting the blank obtained under the same conditions in the absence of protein The CD measurement of HpSDH ... antibacterial activity Experimental procedures Materials H pylori strains SS1 and ATCC 43504 were obtained from Shanghai Institute of Digestive Disease (Shanghai, China) E coli host strain BL21(DE3) ... meningitidis, M jannaschii, Archaeoglobus fulgidus, and Mycobacterium tuberculosis SDH, respectively To obtain the high level of protein production, we reduced the amount of isopropyl thio-b-d-galactoside...
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Tài liệu Báo cáo khoa học: Biochemical characterization of human 3-methylglutaconyl-CoA hydratase and its role in leucine metabolism docx
... 3-Methylglutaconyl-CoA hydratase 3-Hydroxy-3-methylglutaryl-CoA lyase 2.6.1.42 1.2.4.4 BCAT1 BCKDHA BCKDHB DBT DLD IVD MCCC1 AUH HMGCL 113520 608348 248611 248610 246900 607036 609010 250950 246450 3-Methylglutaric ... short-chain substrates was predicted [18] The first pure preparation of a 3-MG-CoA hydratase was obtained from the bacterium Acinetobacter sp which aerobically grows on isovalerate as sole carbon ... CoA-substrates (R,S)-HMG-CoA, crotonyl-CoA, 3-methylcrotonyl-CoA and 3-hydroxybutyryl-CoA were obtained from SigmaAldrich Assay of 3-MG-CoA hydratase The assay contained in a total volume of 25...
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Tài liệu Báo cáo khoa học: Biochemical characterization of Bacillus subtilis type II isopentenyl diphosphate isomerase, and phylogenetic distribution of isoprenoid biosynthesis pathways doc
... Germany) FMN was obtained from Sigma (Steinheim, Germany) Cloning and expression of the idi-2 gene from B subtilis A DNA segment extending from bp position 184 997– 186 043 of the B subtilis chromosome ... isomerase from Bacillus subtilis has been elucidated by X-ray crystallography [29] This paper reports on the biochemical properties of the recombinant enzyme from B subtilis Phylogenetic patterns ... Cloning and expression of the idi-2 gene from B subtilis An open reading frame (Acc no P50740) extending from bp position 184 997–186 043 on the B subtilis chromosome with similarity to the idi-2...
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Tài liệu Báo cáo khoa học: Biochemical and molecular characterization of hazelnut (Corylus avellana) seed lipoxygenases pdf
... bp) of the isolated CaLOX2 A simple hybridization pattern was obtained with both the probes Figure shows the hybridization signals obtained with the 836 bp fragment corresponding to the 3¢ portion ... previously reported [9] LOX activity was assayed in total protein samples and in microsomal fraction obtained by centrifugation of total protein samples at 100 000 g for 30 The pellet (microsomal fraction) ... conserved region (2103–3039) of the soybean LOX3 gene [16] An amplification product of about 700 bp was obtained in the above described conditions The amplified DNA was cloned and sequenced The results...
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Báo cáo khoa học: Biochemical characterization of USP7 reveals post-translational modification sites and structural requirements for substrate processing and subcellular localization pptx
... In addition, an average of mg USP7 208-560 per litre of Escherichia coli fermentation broth was obtained from the soluble cell fraction The recombinant proteins were purified to homogeneity (‡ 90%) ... techniques used, monomers or dimers of the enzyme were detected in vitro, whereas in cells evidence was obtained pointing to oligomerization events Deletion of the N- and C-terminal domains of USP7 affected ... ligase but not by MDM2 [12] Our findings indicate that USP7 could exist as a dimer in cells The data obtained with purified enzyme is, however, contradictory, suggesting that further cellular components...
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