bla m fusion construct and its incorporation into virus like particles vlps
Báo cáo khoa học: "Development of a RVFV ELISA that can distinguish infected from vaccinated animals" pps
... entire biomedical community peer reviewed and published immediately upon acceptance cited in PubMed and archived on PubMed Central yours — you keep the copyright BioMedcentral Submit your manuscript ... rocking at room temperature for hour Mix was allowed to settle in a chromatography column and flow through was collected Column was washed twice with ml of Buffer C (8 M urea, 0.1 M sodium phosphate ... study and the molecular cloning AKM performed the cloning, gene expression, purification, immunoassays and drafted the manuscript STN conceived of the study and participated in its design and coordination...
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Báo cáo khoa học: Molecular cloning, expression and characterization of protein disulfide isomerase from Conus marmoreus pdf
... performed in 0.2 M sodium phosphate buffer (pH 7.5) containing mM GSH, 30 lM insulin, 120 lM NADPH, 0.5 units of glutathione reductase, mM EDTA, and 0.7 lM PDI The absorbance decrease at 340 nm was ... was monitored b Refolding of the reduced RNase A (final concentration 8.4 lM) was carried out in the refolding buffer (0.1 M Tris ⁄ Cl, pH 8.0, 0.2 mM GSSG, mM GSH, mM EDTA, 4.5 mM cCMP) and catalyzed ... mM EDTA, mM MgCl2, 20 mM NaCl, mM GSSG, mM GSH) and catalyzed by different concentrations of PDI In the refolding reaction mixture, the final concentration of reduced lysozyme was 10 lM The refolding...
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Báo cáo khoa học: Cloning, expression and interaction of human T-cell receptors with the bacterial superantigen SSA ppt
... and immunoblot SSA–iodoacetamide (SSAia), with the free Cys blocked, was purified as a monomer by S-75 column (Amersham Pharmacia Biotech AB) with 50 mM MES, pH 6, 150 mM NaCl SDS/PAGE and immunoblotting ... performed using a size exclusion Superdex 75 column (Amersham Pharmacia Biotech AB) equilibrated with 50 mM Tris, pH 7.5, 150 mM NaCl and finally with a Mono-S cation exchange column (Amersham Pharmacia ... same buffer, and finally with 100 mM Tris/HCl, pH 7.5, mM EDTA and mM DTT Inclusion bodies were then solubilized in M urea, 100 mM Tris/HCl, pH 7.5, 10 mM EDTA and mM DTT Concentration of solubilized...
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Báo cáo khoa học: Molecular cloning, expression and characterization of cDNA encoding cis-prenyltransferases from Hevea brasiliensis A key factor participating in natural rubber biosynthesis pdf
... was performed by the modified method of Tangpakdee et al [16] The reaction mixture contained, in a final volume of 0.2 mL, 50 mM Tris/HCl buffer (pH 7.4), 30 mM KCl, mM MgCl2, lM ZnCl2, mM dithiothreitol, ... dithiothreitol, 20 mM KF, 0.1 mM deoxycholate, 0.5 mg of WBP, 15 lM farnesyl diphosphate, 50 lM [1-14C]IPP (2.15 GBqÆmmol)1, Amersham Pharmacia Biotech), and a suitable amount of cell-free homogenate or ... transformation of E coli BL21(DE3), and mL of an overnight culture of the transformant in Luria–Bertani medium containing 50 lgÆmL)1 ampicillin was inoculated into 200 mL of M9 YG medium [29] containing...
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Báo cáo khoa học: Sulfation of hydroxychlorobiphenyls Molecular cloning, expression, and functional characterization of zebrafish SULT1 sulfotransferases docx
... substrates Data are given as mean ± SD from three experiments SULT1 ST1 SULT1 ST2 Substrate Km (lM) Vmax (nmolÆmin)1Æmg)1) Vmax/Km Km (lM) Vmax (nmolÆmin)1Æmg)1) Vmax/Km 3-Chloro-4-biphenylol 3,3¢,5,5¢-Tetrachloro-4,4¢-biphenyldiol ... [13] Ham’s F-12 nutrient mixture, Leibivitz’s L-15 medium, Dulbecco’s modified Eagle’s medium, minimum essential medium, and fetal bovine serum were from Life Technologies Trout serum was from East ... replacing magnesium sulfate with magnesium chloride) minimum essential medium for h, were labeled with 0.2 mL aliquots of the same medium containing [35S]sulfate (0.25 mCiÆmL)1), and 100 lM 3-chloro-4-biphenylol...
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Báo cáo khoa học: Cloning, expression and characterization of a new aspartate aminotransferase from Bacillus subtilis B3 docx
... 1.0–27 mm l-glutamate with a fixed concentration of mm oxaloaceLÀglu ) and in the range 0.5–20 mm oxaloacetate tate (for Km with a fixed concentration of 12 mm l-glutamate OAA (for Km ) Km and Vmax ... Kuramitsu S (1996) An aspartate aminotransferase from an extremely thermophilic bacterium, Thermus thermophilus HB8 J Biochem 119, 135–144 Sung MH, Tanizawa K, Tanaka H, Kuramitsu S, Kagamiyama ... concentrations: ampicillin (Amp), 100 lgÆmL)1; kanamycin (Km), 50 lgÆmL)1 DNA manipulation and transformation The isolation and manipulation of recombinant DNA were performed using standard techniques...
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Báo cáo Y học: Trehalose-phosphate synthase of Mycobacterium tuberculosis Cloning, expression and properties of the recombinant enzyme pdf
... tuberculosis (Mt) OtsA predicted amino acid sequence with those of homologous ORFs from M avium (Ma), M smegmatis (Ms) and E coli (Ec) The M avium and M smegmatis homologs were identified by searching ... heating and the following components were added: 0.15 mM NADH, 0.25 mM phosphoenolpyruvate, mM MgCl2, and lg each of pyruvate kinase and lactate dehydrogenase in a final volume of 200 lL 50 mM Hepes ... recombinant enzyme have been determined, and are compared to the properties of the TPS purified from M smegmatis This expression system should provide sufficient amounts of recombinant TPS for complete...
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Báo cáo khoa học: Trehalose synthase of Mycobacterium smegmatis Purification, cloning, expression, and properties of the enzyme ppt
... maltose was estimated to be 10 mM and the Vmax for maltose was determined as 16 nmolÆmin)1 A similar experiment using trehalose as the substrate showed that the formation of maltose increased ... nickel column The TIGR unfinished M smegmatis genome sequence was screened using the TBLASTN program for DNA sequences corresponding to the amino acid sequences obtained from purified M smegmatis TreS ... ion column and after thorough washing in 10 mm imidazole, the column was eluted batchwise with various concentrations of imidazole Most of the activity was eluted in 100 mM imidazole, and as shown...
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Báo cáo khoa học: Cloning, expression and characterization of a family-74 xyloglucanase from Thermobifida fusca pptx
... three volumes of 0.4 M NH2SO4 + mM NaCl + mM Kpi, pH 6, and eluted with mM Kpi, pH The purest fractions, as determined by SDS/PAGE, were combined and loaded on a Q-Sepharose Fast Flow (Pharmacia ... nature and presence of a CBM varies among the family-74 proteins: four of 13 family-74 genes code for a family-2 CBM; three have a family-1 fungal CBM; one has a family-3 CBM; and five have no CBM ... concentrations from to 60 lgÆmL)1 and 0.375 pmolÆmL)1 enzyme The bicinchoninate assay [17] was used to quantify the reducing sugars produced Michaelis constant (Km) and maximal rate (Vmax) values were...
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Báo cáo khoa học: Gene cloning, expression and characterization of avian cathelicidin orthologs, Cc-CATHs, fromCoturnix coturnix pdf
... Fungi Candida albicans ATCC 2002 Candida glabrata (IS 0902) Slime mold 1580 Cc-CATH2 (0 mM NaCl) Cc-CATH2 (100 mM NaCl) Cc-CATH3 (0 mM NaCl) Cc-CATH3 (100 mM NaCl) LL-37 (0 mM NaCl) Amp Kana ... phase at 37 °C and suspended to · 105 colony-forming unitsÆmL)1 in Mueller– Hinton (MH) broth with and without 100 mm of NaCl The peptides in the presence and absence of 100 mm NaCl were subjected ... essential gene-encoded mammalian antibiotics J Mol Med 80, 549–561 Zanetti M (2005) The role of cathelicidins in the innate host defenses of mammals Curr Issues Mol Biol 7, 179– 196 Ramanathan B, Davis...
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Báo cáo Y học: Cloning, expression and characterization of a gene encoding nitroalkane-oxidizing enzyme from Streptomyces ansochromogenes pot
... Control MnCl2 (1 mM) CuSO4 (1 mM) CaCl2 (1 mM) MgSO4 (1 mM) NAD (2 mM) Cystine (2 mM) 100 117.8 16.9 82.0 89.8 92.5 45.8 2-Mercaptoethanol (2 mM) GSH (2 mM) NADH (2 mM) EDTA (2 mM) NBT (5 mM) SOD ... carbonyl compound formed in the reaction solution was further confirmed to be acetone by its mass spectrum, which displayed fragments of m/ z 58 (M+ ) and m/ z 43 consistent with those of acetone standard ... Restriction enzymes and T4 DNA ligase were purchased from Boehringer-Mannheim and Sino-American Biotechnology Company (Luoyang, Chi1 na) DIG labeling and detection kits (Boehringer-Mannheim) were used...
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Báo cáo y học: "Cloning, expression and characterization of gE protein of Duck plague virus" doc
... rpm/min for 10 min, and lysed in 20 ml 20 mM Tris-HCl pH 8.0 containing 1.0 mg/ml lysozyme at -20°C overnight The cell lysate was clarified by centrifugation at 10,000 rpm/min for 20 at 4°C and ... The immunofluorescence studies showed that gE mainly localized in the cytoplasm, and DPV gE might share similar functions with its HSV-1, VZV-1, and PRV homolog gE The real time PCR, RT-PCR, and ... nuclear membrane, which would fuse with the membrane of the endoplasmic reticulum The naked nucleocapsids were released into the cytosol, and they became enveloped during budding into cytosolic membraneous...
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Cloning, expression and characterization of novel helicobacter pylori differentiating antigen heat shock protein 20
... mass Spectrometry ML: maximum likelihood MS: mass spectrometry MW: molecular weight viii NUD: non-ulcer dyspepsia OD: optical density OMP: outer membrane protein OPD: O-phenylenediamine dihydrochloride ... HSP60 and its homologue in human by both Kansau & Labigne (1996) and Yamaguchi et al (2000) It was speculated that H pylori HSP60 might be responsible for molecular mimicry causing autoimmune ... (1999) examined the Ks, Ka and Ka/Ks of vacA alleles m1 and m2 The results showed that the greatest difference was present in the comparison between m1 and m2 but not within m1 or m2 Since the...
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Cloning, expression and characterization of oxysterol binding protein homologue 7 (OSH7) in yeast saccharomyces cerevisiae
... HOMEOSTASIS Sterols are important membrane components of all known eukaryotic organisms and play essential roles in modulating membrane fluidity and permeability In mammalian cells, the predominant ... function in membranes of animal cell, and because it is the raw material for the manufacture of steroid hormones and bile acids The very property that makes it useful in cell membrane, namely its absolute ... in arteries 41 MATERIALS AND METHODS 2.1 MEDIA, REAGENTS, STRAINS AND PLASMIDS Most chemicals were from Sigma (St Louis, MO, USA) unless otherwise stated Luria-Broth (LB) medium used for bacterial...
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Tài liệu Báo cáo khoa học: Molecular cloning, recombinant expression and IgE-binding epitope of x-5 gliadin, a major allergen in wheat-dependent exercise-induced anaphylaxis ppt
... difluoride membrane (Immobilon-P, Millipore, Billerica, MA, USA) and blocked with 5% skim milk in TBST (50 mm Tris-bufferd, sa1ine % Tween 20, pH 4.7) The membrane was washed three times with TBST and ... and QQFHQQQ, are not common but might be important epitopes for the development of allergic symptoms in WDEIA We cloned and determined the nucleotide sequence of two x-5 gliadin genes, x-5a and ... x-5 gliadin fraction from T aestuvum Butte was resolved into three peaks of molecular mass 49 085, 50 300, and 51 500 by MALDI-TOF MS [9] However the calculated molecular mass (50 900) of x-5a...
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Tài liệu Báo cáo khoa học: Cloning, characterization and expression analysis of interleukin-10 from the common carp, Cyprinus carpio L. docx
... Spotted green pufferfish Human Cat Rat Mouse Virus IL-10 homologues Human Epstein–Barr virus IL-10 Equine herpes virus type IL-10 Human cytomegalovirus Human IL-10 family members IL-19 IL-20 IL-22 ... using PCR primers based on the conserved regions of mammalian IL-10 homologues and the torafugu IL-10 genomic sequence, we isolated a clone of 440 bp (resembling mammalian IL-10) from carp head ... to that of torafugu and mammalian counterparts Compared with other family members containing the IL-10 gene, human IL-19 and IL-24 encode longer polypeptides of 215 and 206 amino acids, respectively;...
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Báo cáo khoa học: Molecular cloning, expression analysis and functional confirmation of ecdysone receptor and ultraspiracle from the Colorado potato beetle Leptinotarsa decemlineata pdf
... deionized formamide, 9.2% v ⁄ v glycerol, 0.18 mgÆmL)1 bromophenol blue, 0.18 mgÆmL)1 xylene cyanole and 0.4 mm EDTA) was denatured at 65 °C for 15 and immediately transferred on ice This RNA sample ... on 1% agarose gel (20 mm Mops, mm sodium acetate, 0.5 mm EDTA and 18% deionized formaldehyde) and hydrolyzed in 50 mm NaOH solution for 25 After neutralization with 0.2 m acetic acid solution ... ecdysteroid receptor homologue from the ixodid tick Amblyomma americanum (L.) Insect Biochem Mol Biol 27, 945–962 28 Guo X, Xu Q, Harmon MA, Jin X, Laudet V, Mangelsdorf DJ & Palmer MJ (1998) Isolation...
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Báo cáo Y học: Molecular cloning, bacterial expression and properties of Rab31 and Rab32 docx
... Immobilon-P membrane for blotting proteins, HAWP ®lters (0.45 lm, 25 mm) and Centricon YM-10 ®lters were from Millipore RPMI 1640 medium, foetal bovine serum, L-glutamine, T4 DNA ligase, Taq DNA polymerase, ... times, triplicate 10 lL samples of the incubation mixture were diluted into 100 lL of wash buffer [100 mM KCl, 20 mM Hepes (pH 7.5), 0.5 mM MgCl2] and immediately applied to HAWP ®lters in a Millipore ... Biochem Biophys Res Commun 195, 1081± 1088 Pereira-Leal, J.B & Seabra, M. C (2000) The mammalian Rab family of small GTPases: de®nition of family and subfamily sequence motifs suggests a mechanism...
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Báo cáo khoa học: Bovine tryptases cDNA cloning, tissue specific expression and characterization of the lung isoform ppt
... digested and ligated genomic DNA sample (10 lL), using 20 pmoles of each primer (see below) and U of the ÔElongase enzyme mixÕ (Gibco) in 60 mM Tris sulfate pH 9.1, 18 mM ammonium sulfate, mM magnesium ... Concentration mM mM mM 0.1 mgÆmL)1 0.1 mgÆmL)1 0.1 mgÆmL)1 100 0 96 100 35 Discussion In previous studies, we reported isolation of tryptase (then named BLCT) from bovine liver capsule and its characterization ... (Perkin Elmer, USA), 200 mM dNTPs, 1.5 mM MgCl2, 10 mM Tris/HCl, pH 8.3, 50 mM KCl (50 lL final volume) All PCR products were sizefractionated by agarose gel electrophoresis and the bands eluted,...
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Báo cáo khoa học: Identification of a novel thyroid hormone-sulfating cytosolic sulfotransferase, SULT1 ST5, from zebrafish Molecular cloning, expression, characterization and ontogenic study ppt
... determined is lower than the detection limit (estimated to be 0.01 nmolÆmin)1Æmg protein)1) Endogenous compounds Specific activity (nmolÆmin)1Æmg)1) Xenobiotic compounds Specific activity (nmolÆmin)1Æmg)1) ... room temperature, cells were collected by centrifugation and homogenized in 25 mL ice-cold lysis buffer (10 mm Tris ⁄ HCl, pH 8.0, 150 mm NaCl, mm EDTA) using an Aminco French Press Twenty microliters ... 2.5 mL of glutathione Sepharose, and the bound GST fusion protein was treated with mL of a thrombin digestion buffer (50 mm Tris ⁄ HCl, pH 8.0, 150 mm NaCl, and 2.5 mm CaCl2) containing unitÆmL)1...
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