isolation and characterization of alkaline phosphatase

... polysaccharide One milligram of the partially purified polymer contained 309.7 µg of protein, 160.2 µg of neutral sugar, 58.5 µg of hexosamine, 30.8 µg of uronic acid and 3.8 µg of phosphorus Constituent ... concentration of ppm to estimate the effect of pH After min, A660 was measured and the extent of flocculation was calculated Pronase digestion of extracellular polymer Forty milligrams of the extracellular ... as follows: 500 g of soil was washed times with distilled water and suspended in 1,000 ml of water The mixture was allowed to stand for 10min and the supernatant was collected and used for the...

Ngày tải lên: 05/09/2013, 09:08

... for cDNA single strand synthesis, 600 ng of RNA, 0.5 lg of oligo(dT) primer and 10 mm of dNTPs were incubated for at 65 °C Subsequently, lL of the first strand buffer X and lL of dithiothreitol ... III and P IV isoforms from the eluate P III ⁄ P IV (Fig 1C) Isoelectric focusing gave pI of 5.8, 5.7, 5.5 and 5.2 for the four isoforms respectively Secondary structure of the pulchellin isoforms ... mixture of other isoforms (B) Gel visualization of proteins eluted Lane 1, molecular weight markers Numbers on the left indicate the Mr values of the standards in thousands Lanes 2–9, lg of peak...

Ngày tải lên: 18/02/2014, 16:20

... of 50 nmÆmin)1, band-width of 1.0 nm, and a resolution of 0.1 nm in the temperature range between 10 °C and 70 °C, after preincubation for 10 at each temperature Three runs were accumulated and ... of 0.6 lM The data were fitted to Michaelis–Menten curves Squares and dotted line: data and fitting with PHI/PHO ratio of 0.5 Triangles and continuous line: data and fitting with PHI/PHO ratio of ... Asterisks and broken line: data and fitting with PHI/PHO ratio of hypothesis and therefore a direct involvement of PHI in electron transfer This conclusion is consistent with the very low degree of identity...

Ngày tải lên: 21/02/2014, 00:20

... intense band of 61 kDa (NrfA) and a band of weak intensity of 19 kDa (NrfH), confirming its hetero-oligomeric nature (Fig 1, lane 1) However, in the absence of boiling (Fig 1A, lanes and 4) high ... and 4) high molecular mass bands of approximately 110 kDa and > 200 kDa were visible, as well as a faint band at 37 kDa, suggesting the presence of dimers All of the bands stained positively for ... suggests that the bands at approximately 110 and 37 kDa correspond to strongly attached dimers of both NrfA (a2) and NrfH (b2), respectively The presence of dithiothreitol (Fig 1A, lanes and 2) did not...

Ngày tải lên: 21/02/2014, 00:20

... RESULTS Cloning and sequences of Ch-Prx1 cDNA and the Ch-Prx1 gene In order to complete the sequence data for this new protein, and to be able to make a thorough characterization of its biochemical ... BLAST of peptide sequences of 21 kDa proteins in databases Tryptic peptides were puri®ed by RP-HPLC and some of them were totally or partially analyzed by Edman sequencing (ááá) and/ or by MALDI-TOF ... BAS1 protein of Brassica, spinach, barley and A thaliana, PR1 of Phaseolus and MHF of A thaliana These proteins belong to the 2Cys-Prx subfamily All plant 2Cys-Prx proteins, except BAS1 of barley,...

Ngày tải lên: 08/03/2014, 16:20

... presequence of 19 amino acids The mature protein would have a molecular mass of 41.7 kDa and a pI of 6.34 The YedO protein from E coli and the d-CDes from Arabidopsis showed an overall identity of 36% and ... d-CDes mRNA and protein levels in older plants, during the light phase in a diurnal light ⁄ dark rhythm and under sulfate limitation Results In silico characterization and isolation of the Arabidopsis ... protein has a predicted molecular mass of 43.9 kDa and a pI of 7.2 It contains relatively high amounts of the sulfur amino acids cysteine (four residues) and methionine (10 residues) According...

Ngày tải lên: 16/03/2014, 18:20

... respectively, and the resultant VNARs evaluated for retention of binding affinity Material and methods Equipment and reagents Restriction enzymes and Vent DNA polymerase were purchased from New England ... possibility of inter- and intra CDR disulphide cross-links These strategies are summarized in Fig 1B and Table 1, and details of the library construction are given in greater detail in the Materials and ... possibility of disulphide bond formation genetically by replacement of residues Cys22 and Cys82 with alanine and valine, respectively, to give a cysteine minus mutant (12F-11DCys) Use of alanine and...

Ngày tải lên: 17/03/2014, 10:20

... site molecules Y80, Y123 and E177 are shown in green and the position of the two mutated amino acids, F108 and L151, are shown in blue between the aromatic rings of Tyr80 and Tyr123 close to the ... diagram of wild-type RTA structure, and the positions of the altered amino acids of the double mutant, F108 and L151, within the structure are indicated Discussion RTA is the catalytic polypeptide of ... Utilisation of the budding yeast Saccharomyces cerevisiae for the generation and isolation of non-lethal ricin A chain variants Yeast 22, 1287–1297 16 Gemmill TR & Trimble RB (1999) Overview of N- and...

Ngày tải lên: 23/03/2014, 07:20

... Switzerland), thermolysin, chymotrypsin or elastase (Sigma, St Louis, MO, USA)] and Cloning of the bovine MUC15 cDNA by PCR with degenerate primers Isolation of total RNA from the mammary gland of ... PCR screening of bovine and human cDNA libraries showed the presence of MUC15 mRNA in human breast, bovine mammary gland, bovine lymph nodes and lungs of both species (Table 1) Of the identified ... part of the coding sequence was then cloned and sequenced using PCR as described above with 0.4 lM and lM of the specific P7 primer and degenerate primers (P4-P6), respectively The cDNA sequence of...

Ngày tải lên: 24/03/2014, 04:21

... of degenerate and specific primers and by rescreening the cDNA libraries The partial clones of XHIVEP2 and -3 covered 5.9 and 3.2 kb of the respective 3¢ ends and included 3¢-UTRs and poly(A)-tails ... upper strands with [32P]ATP[cP] and T4 polynucleotide kinase The labeled oligomers were annealed by heating to 90 °C an equimolar mixture of the upper and lower strands in reaction buffer and cooling ... -2 and -3) The Xenopus XHIVEPs are characterized with respect to temporal and spatial expression, nuclear import/export activity and DNA binding specificity Materials and methods Isolation and...

Ngày tải lên: 30/03/2014, 13:20

... Domains of E9 and locations of mutations responsible for drug resistance Domains of E9 and locations of mutations responsible for drug resistance The location of the putative exonuclease and polymerase ... streptomycin and non-essential amino acids Vero Cells were obtained from ATCC and were maintained in MEM with Earl's salts and the addition of 10% FBS and standard concentrations of L-glutamine, ... Mapping and marker rescue of recombinant viruses A Map of PCR fragments in the E9L region that were used for marker rescue mapping experiments and reconstruction of resistant viruses B Results of...

Ngày tải lên: 20/06/2014, 01:20

... the G and P serotype specificities of the YK-1 strain, the sequences of the genes encoding both VP7 and VP4 were determined and compared with those of representatives of established G and P serotypes ... rotavirus RNA segment 11, and (c) schematic diagram of the sequences of gene segment 11 Arrows indicate segment 11 of RRV and YK-1, or rearranged segment 11 of variant 311 The YK-1 and its variant clone ... Electrophoretic study of the genome of human rotaviruses from Rio de Janeiro, Sao Paulo and Belem, Brazil J of Hygiene 1983, 90:117-125 Palombo EA, Bishop RF: Genetic and antigenic characterization of a serotype...

Ngày tải lên: 20/06/2014, 01:20

... mM EDTA, 1.5 M NaCl), in the presence of 0.1 mg of proteinase K and mg of lysozyme and incubated at 37°C for 30 Lysis was completed by adding ml of 20% SDS and incubated for h at 65°C with shaking ... 65 and 85% of the maximum activity at the temperature range of 35°C and 55°C respectively Thermal stability of the purified AS-protease was estimated at Figure Multiple sequence alignment of ... Waschkowitz T, Rockstroh S, Daniel R (2009) Isolation and characterization of metalloproteases with a novel domain structure by construction and screening of metagenomic libraries Appl Environ Microbiol...

Ngày tải lên: 21/06/2014, 05:20

... eht fo esahp suoeuqa reppu eht no detcelloc saw AND lairetcaB nim rof 000,6 ta degufirtnec dna nim rof sllec lairetcab gniliob yb detcartxe neht saw AND lairetcab ;retaw dellitsid elbuod dezilirets ... 817( rof evitisop ,9~4 senal ;)pb 001,1( rof evitisop ,3 dna senal ;noitcefni laccocotperts rof evitagen ,1 enal ;lortnoc evitagen ,N enal ;)pb 003 ,M4040 ( lortnoc evitisop ,P senal ;reddal AND ... enilav ,esapil rof snoitcaer evitagen dah setalosi llA nispyrtomyhc -α dna esatahpsohp enilakla rof snoitcaer evitisop dah fo setalosi 81 dna esadinimasoculg-β-lyteca -N dna esaretse rof snoitcaer...

Ngày tải lên: 07/08/2014, 18:21

... condition C, E and G: Undifferentiation; D, F and H: Neuronal differentiation Nestin, Tuj-1 and NF160 were green GFAP and MAP2 were red Scale bars = 50 μm Isolation and characterization of canine ... (Figs 4E and F) Discussion Isolation and characterization of stem cells derived from various tissues and sources have been one very critical issue for stem cell therapy [12,28,33] The purpose of this ... stem cells from dogs The characterization conditions and basic settings for the application of gene delivery were also investigated Materials and Methods Cell isolation and culture Canine umbilical...

Ngày tải lên: 07/08/2014, 23:22

... letters showed site of H274Y and N294S substitutions (H275Y and N295S in N1 influenza numbering) cough, fever and serous nasal discharge The course of the disease was short (72 h) and responsive well ... Details 1Department of Virology, Faculty of Veterinary Medicine, Beni-Suef University, Beni-Suef 62511, Egypt, 2Division of Virology, Department of Microbiology, College of Medicine and Medical Sciences, ... Glisic S, Perovic V, Köhler H: Characterization of conserved properties of hemagglutinin of H5N1 and human influenza viruses: possible consequences for therapy and infection control BMC Struct...

Ngày tải lên: 10/08/2014, 05:21

... temperature of the heat treatment to 70°C, band X faded, whereas the intensity of band Y increased With further increases in temperature, band Y faded, whereas the intensity of band Z increased; ... only band Z was observed The sizes of proteins in bands X, Y, and Z were estimated to be 370 kDa, 105 kDa, and 50 kDa, respectively, compared to size markers To identify proteins of these bands, ... surface of the algal cell wall (TEM, upper panel) and released particles (SEM, lower panel) of Chlorella variabilis virus CvV-BW1 is on the left (A and C) and CvV-BW3 is on the right (B and D)...

Ngày tải lên: 12/08/2014, 01:21

... sativus HT performed the isolation and heterologous expression of the HTMT encoding gene from R sativus and characterization of the enzymatic properties of recom- Page of 10 (page number not for ... respectively, and the products were quantified using the peak area To calibrate the concentrations of the products, gas and liquid standards of CH3I, DMS, and CH3SCN were prepared The detection limits of ... from R sativus and expressed in Escherichia coli This paper reports the emission profiles of methyl halides from some plants and the characterization of the enzymatic properties of recombinant...

Ngày tải lên: 12/08/2014, 03:21

... DNAstar software package Sequence analysis based on the complete ORF2 sequence of the isolate in this study and other 10 genotype HEV isolates in China, using DNAstar software package Page of (page ... participated in the planning of the project XGH was the leader of the project WZ, SXY, QS, and TLS collected all samples and performed the RT-PCR experiments for HEV RNA WZ and SXY did the phylogenetic ... Bezanilla JC, Toth TE, Meng XJ: Identification of genotype hepatitis E virus (HEV) in serum and faecal samples from pigs in Thailand and Mexico, where genotype and HEV strains are prevalent in the respective...

Ngày tải lên: 12/08/2014, 04:21

... identification and characterization of MPs in the sputa of CF patients Patients and Methods Study patients The study was approved by, and performed in accordance with, the ethical standards of our institutional ... monitoring of CF and other respiratory diseases, beside for the study of disease pathogenesis and its treatment Measurement of these components is largely sophisticated and quantifiable, and the ... CF and PCD sputum) Statistical significance of differences between acute and stable groups of CF patients was evaluated by a two-tailed unpaired Student's t-test To compare the number of MPs and...

Ngày tải lên: 12/08/2014, 11:22