fi cation and detection of nucleic acids puri fi ed from dbs

... versatility of the electrode was tested Cyclic voltammetry was used for the detection of two nucleic acids samples mentioned above and the basic electrochemical behaviour of nucleic acids at the ... treated electrochemical data and participated in preparation of the manuscript VA participated in the design of the study and performed the analysis of the data RK conceived of the study, and ... considered as a suitable tool for detection of nucleic acids We show the successful application of modern nano-technologies not only for detecting of nucleic acids but also for distinguishing of all...

Ngày tải lên: 21/06/2014, 03:20

... representation of the purification of sense p185-ssDNA from biotinylated p185 dsDNA 139 Figure 4.4 Agarose gel electrophoretic image of the double-stranded (Lane 2) and single-stranded DNA ... enabled the detection of hybridization Such procedure eliminated the use of external labels Figure 1.2 shows the basic principle of label-free nucleic acids detection based on the monitoring of ... Hybridization and Detection of Nucleic Acids 1.1.1 Basic of nucleic acid hybridization The nucleic acid is a polymer consists of monomers called nucleotides Each nucleotide composes of a nitrogenous...

Ngày tải lên: 12/09/2015, 09:28

... Turtle liver GST puri cation The puri cation procedure was developed using liver extracts from control turtles but also used for puri cation of GSTs from anoxic liver Four milliliters of crude supernatant ... was puri ed with a combination of three chromatography methods: dye ligand, gel filtration and ion exchange The puri cation scheme developed for turtle GSTs resulted in final specific activities of ... profiles from Sephacryl Table Puri cation of GST from liver of control and anoxic turtles Enzyme activity was assayed using optimal CDNB and GSH concentrations Results are from a single puri cation...

Ngày tải lên: 23/03/2014, 15:20

... Expression and puri cation of recombinant proteins DNA fragments encoding predicted mature regions of recombinant proteins were ampli ed by PCR using each specific primer set Ampli ed PCR products ... sequence analysis of cDNA clones H longicornis salivary gland mRNA was isolated from 10 pairs of glands of ticks at three distinct feeding stages (unfed, slow feeding, and rapid feeding) by using ... Identi cation of tick thrombin inhibitors, madanin and (Eur J Biochem 270) 1929 unfed, slow feeding, and rapid feeding A total of 1889 cDNA clones were picked from three cDNA libraries, and their...

Ngày tải lên: 31/03/2014, 01:20

... shown) To confirm that the cDNA cloned above was that of the GAT puri ed in this study, we screened a cDNA library (5 · 106 primary plaques) prepared from the mixture of stressed and unstressed leaves ... clone of the GAT, genespecific primers were designed from the sequence of watermelon EST AI563351 and used for 5¢- and 3¢-RACE The cloned cDNA encoded a protein 5356 K Takahara et al composed of ... Puri cation of GAT from wild watermelon leaves To characterize the GAT in detail, it was puri ed from wild watermelon leaves (Table 1) The puri cation procedure was developed by taking advantage of the...

Ngày tải lên: 20/02/2014, 03:20

... Results Puri cation of lectin from the hemolymph of P jacquemontii A column profile depicting puri cation of lectin from the hemolymph of P jacquemontii by affinity chromatography on BSM-coupled Sepharose ... Clari ed serum (20 mL) was applied and on elution with EDTA yielded 1.0 mg of pure lectin The specific activity of puri ed lectin increased about 2000-fold from 196 (crude hemolymph) to 409 600 of ... evaluation of the literature revealed that puri cation of lectin from the hemolymph of crustaceans was most successful by affinity chromatography as it gave a higher fold of puri cation and percentage of...

Ngày tải lên: 21/02/2014, 00:20

... was detected, tetrazolium was reduced, nicotinamide was required for growth and potassium tellurite inhibited growth Thus, strain 10d was identi ed as a species of Bordetella Puri cation and properties ... Classi cation and Identi cation of Microorganism (Hasegawa, T., ed. ), pp 99–160 Japan scientific societies press, Tokyo 16 Johnson, R & Sneath, P.H.A (1973) Taxonomy of Bordetella and related organisms ... All steps of the enzyme puri cation were carried out at 0–4 °C All centrifugations were at 20 000 g and °C for 10 A wet weight of 30 g of Bordetella sp strain 10d cells were obtained from a 4.8-L...

Ngày tải lên: 21/02/2014, 01:21

... Enzyme puri cation All puri cation steps were carried out under strictly anoxic conditions under an atmosphere of N2/H2 (95 : 5, v/v) at 18 °C Cells were lysed by sonication and then centrifuged ... b and two Table N-Terminal sequences of the polypeptides of the puri ed enzyme N-Terminal sequences were either obtained by Edman degradation (column 1) or derived from the genome sequence of ... the reduced state Any enzyme molecules that contained oxidized heme groups could be rapidly reduced by sodium dithionite Addition of DMN to the reduced enzyme resulted in rapid oxidation of the...

Ngày tải lên: 21/02/2014, 03:20

... phylogenetic analysis of fish hatching enzyme genes, hatching enzyme originally consisted of a single type of enzyme, and HCE and LCE were produced by duplication and diversi cation of the gene [16] ... ZHE1 gene, and not the ZHE2 gene, is predominantly expressed in zebrafish embryos Puri cation of ZHE The hatching liquid (i.e culture medium after embryos hatched out) was used to purify ZHE First, ... sequence from the N-terminus of mature protein portion deduced from ZHE1 cDNA, 5936 Fig Puri cation of the hatching enzyme of zebrafish (A) Superdex 75 10 ⁄ 300 GL column chromatogram of hatching...

Ngày tải lên: 07/03/2014, 04:20

... our puri cation was hampered by the limited amount of starting material available Therefore, fractions VXH-I–VXH-V from the first ion exchange chromatography were pooled, and rechromatographed ... polypeptide composition As many of smaller contaminating polypeptides were observed, apart from bands of the expected mass (22–30 kDa), further puri cation was performed by gel filtration on a Sephacryl ... precursors and are activated in response to wounding of the plant [26] Similar to other known cysteine proteinases from Caricaceae, the deduced protein sequences of VXH-A, -B, -C and -D, and VS-A and...

Ngày tải lên: 07/03/2014, 11:20

... (cytoplasmic marker) and succinate dehydrogenase (cytoplasmic membrane marker) Puri cation of tetrathionate hydrolase All puri cation procedures were performed at °C Washed cells (6 g of wet weight) ... activity of puri ed enzyme The reaction was performed in a thermostat-controlled chamber at 40 °C The reaction was started by addition of protein into mL of preincubated reaction mixture and 500 ... Specific activity (mean ± SD) from two independent experiments Table Puri cation of tetrathionate hydrolase from A caldus One unit (U) is defined as the amount of tetrathionate hydrolase required...

Ngày tải lên: 07/03/2014, 14:20

... Uppsala, Sweden) Results Fig Puri cation of the flavonoid 4¢-OMT through gel filtration Enzymatic activity is expressed as nkatÆmg protein)1 Puri cation of F 4¢-OMT The whole sequence of chromatographic ... activity The different phases of F 4¢-OMT puri cation are presented in Table The enzyme was puri ed 1399-fold, to obtain a final specific activity of 1175 nkatÆmg protein)1 From crude total protein ... were recorded in a glycerol matrix on a VG Prospec (Fisons Instruments, Danvers, NJ, USA) instrument (Cs+ ions of energy of kV) Extraction and puri cation of F 4¢-OMT All the puri cation steps...

Ngày tải lên: 08/03/2014, 08:20

... are defined Fig SDS/PAGE visualized by silver staining of puri ed cod kininogen and wolf fish kininogen, and MALDI-TOF mass spectra of kininogens puri ed by reversed-phase chramotography Ó FEBS ... activity of cod and wolffish kininogen was measured with samples puri ed by reversed phase chromatography Both kininogens were found to inhibit papain and ficin (cysteine proteinases) but neither of ... yield of reversed phase puri ed wolffish kininogen was  850 lg from 200 mL of skin extract We have previously isolated two high molecular mass cysteine proteinase inhibitors from the skin of Atlantic...

Ngày tải lên: 08/03/2014, 23:20

... electrophoresis and N-terminal sequence of the puri ed enzyme (A) Native PAGE of the puri ed enzyme (0.5 lg) in the absence of SDS (B) SDS ⁄ PAGE of the puri ed enzyme (2 lg) and N-terminal sequence of bands ... of these enzymes have not been determined Vignain was puri ed from germinating cotyledons of soybean [3], aleurain was puri ed from barley aleurone layers [4] and caster bean endosperm [5], and ... Boc-Leu-Arg-Arg-4-methylcoumaryl 7-amide (MCA) of the final preparation was 22.9 lmolÆmin)1Æmg)1 About 100 lg of puri ed enzyme was obtained from kg of cotyledons The puri ed enzyme did not bind concanavalin...

Ngày tải lên: 16/03/2014, 04:20

... or puri ed His6-RocF Arginase of H pylori 43504, or puri ed recombinant His6-RocF, was heat-activated at 50 °C for 30 in the presence of various divalent cations, and its activity was measured ... determined that His6-RocF lost Ó FEBS 2004 Characterization of H pylori arginase (Eur J Biochem 271) 1957 Fig pH and temperature optima of puri ed His6-RocF Puri ed His6-RocF was heat-activated with ... Storage of puri ed RocF at °C instead of )20 °C resulted in > 90% loss of activity within week Dialysis of the enzyme also resulted in > 90% loss of enzyme activity (data not shown), as observed with...

Ngày tải lên: 16/03/2014, 16:20

... was puri ed from L E coli culture The molecular mass of the recombinant protein was also analyzed by MS The MALDI-TOF spectrum of puri ed r-Pfen contained three peaks at m/z 25707, 51 383.04 and ... protein puri cation Native enolase from P falciparum strain K1 [17] and strain 3D7 (NCBI: NP_700629) are predicted to contain 446 amino acids The PCR ampli cation of the enolase gene from the ... from thermophilic [12] and sulfate-reducing bacteria [28] The oligomeric state of none of the apicomplexan enolases has been reported so far Kinetic characterization Puri ed r-Pfen was assayed...

Ngày tải lên: 16/03/2014, 18:20

... supernatant was used for further puri cation NTPDase puri cation The solubilized proteins was applied to a column (5 mm · 1.5 cm) filled with Con A/Sepharose 4B and equilibrated with 20 mM start ... ATP/ADPase active protein from puri ed synaptic Ó FEBS 2003 NTPDase1 and NTPDase2 from porcine synaptosomes (Eur J Biochem 270) 3449 Table Puri cation of NTPDase1 and NTPDase2 from porcine brain cortex ... instructions Isolation of NTPDases from synaptosomes Isoelectric focusing of NTPDase1 and NTPDase2 under nondenaturing conditions Synaptosomes were isolated and puri ed by the method of Jones and Matus [23]...

Ngày tải lên: 17/03/2014, 10:20

... lectins puri ed from mulberry seeds are quite different from most of the lectins puri ed from other sources, the subunit structure of mulberry seed lectins are very similar to those of ant egg ... the puri cation steps and over 96% of protein was lost, the puri cation of the proteins was increased after each subsequent puri cation step This low yield may be due to denaturation of the protein ... lengthy puri cation procedure Molecular masses of the lectins and their subunits The molecular masses of the lectins, as determined by gel filtration, were estimated to be 175 000, 120 000 and 89...

Ngày tải lên: 18/03/2014, 01:20

... final concentrations of mM RESULTS Puri cation of VanXYC The vanXYC gene was expressed in E coli JM83 (pAP1) and conventional puri cation of native VanXYC attempted as described in Materials and ... time points and added to 750 lL of cadmium-ninhydrin stock solution; 70 lL of distilled water was added and incubated at 85 °C for The absorbance was measured at 505 nm and quanti ed with free ... included BsaI and EcoRI restriction sites, respectively (underlined) The product was puri ed, digested with BsaI, treated with DNA polymerase I large (Klenow) fragment, puri ed, digested with...

Ngày tải lên: 18/03/2014, 01:20

... identi ed and puri ed from the German cockroach in this study Amino acid sequences obtained from the puri ed GST as well as from cDNA 1798 clones suggested that the enzyme is a member of the ... ions formed from protease-digested peptides The length of these peptides was six to eight amino acid residues The deduced amino acid sequence of one Table Substrate specificities of puri ed cockroach ... deduced sequence of DDS(L ⁄ I)YP(K ⁄ Q), appeared to be closely related to the peptide DDSLYPK identi ed previously in Delta class GSTs of Manduca sexta and D melanogaster [19,24] The deduced...

Ngày tải lên: 23/03/2014, 09:21