searching quanti fi cation and categorization of the identi fi ed proteins

... zffoxp3-F2 and zffoxp3-R2 for foxp3 These primers ampli ed part of the initial predicted sequence which contained the majority of the open reading frames of zebrafish stat6, t-bet and foxp3 to check they ... within the genomic DNA were identi ed, and the amino acid sequences were analysed using blast [26] and fasta [25] This analysis identi ed regions within the zebrafish genome that appeared to code ... sequenced within bony fish, in the Ginbuna crucian carp [27], and STAT6 in mandarin fish [28], whereas Foxp3 has been characterized for the first time in fish The availability of sequenced fish genomes...

Ngày tải lên: 16/02/2014, 09:20

... Department of the Weizmann Institute by a single injection of 150 lg of the puri ed protein, followed by two booster shots of the same amount of protein at 2-week intervals Serum was collected and used ... anti-GroEL Ig [27] The high yield of the UP12-GroEL complex isolation and the ATP-mediated dissociation of the complex strongly support the suggestion that the two stress proteins GroEL and UP12 interact ... GroEL from 15 and 30 lg of the cell lysate without (lanes and 3) or in the presence of ATP (lanes and 4) was detected with anti-UP12 Ig Samples (3 and lg of total proteins) of the cell extract...

Ngày tải lên: 22/02/2014, 07:20

... The mass values of predicted peptides were calculated on the basis of the sequence of goldfish preproprotein On the nano ESI-TOF-MS, a molecular ion peak in the spectrum of the substance eluted ... was identi ed in the goldfish brain The localization of its transcript in the goldfish brain was further investigated 45 °C and 2.5 at 72 °C, and of 30 cycles for 30 s at 94 °C, 30 s at 50 °C and ... sequence of the tridecapeptide, goldfish LPXRFamide peptide-3 subjected to the reversed-phase HPLC puri cation, and the eluate was fractionated every Each isolated substance was then examined by...

Ngày tải lên: 08/03/2014, 16:20

... gene A and B transcripts were determined and comparison of the two sequences showed a 94.1% identity within the coding regions The degree of sequence identity between the Xenopus APLP2-A and APLP2-B ... duplication in the preAPLP family may have resulted in the appearance of the APLP1 and APLP2 gene families The fact that both mammals and Xenopus contain APP and APLP2 proteins suggests that the first ... modi cation in murine APLP2 [24] Furthermore, the transmembrane domains and the cytoplasmic tails of the Xenopus and mammalian proteins are 91% and 100% identical, respectively, including the...

Ngày tải lên: 16/03/2014, 16:20

... analysis showed that the Escherichia coli clones selected induced synthesis of a protein product of the expected size (about 15 kDa) The sequence of the lys gene in the plasmid was checked by sequencing ... consists of 137 amino acids (expected molecular mass of 15.266 kDa) and has a calculated pK value of 8.32 Gene cloning and protein puri cation The lys gene was cloned into plasmid vector pET3a, and the ... antibacterial action of endolysins is determined by the enzyme type, composition of the cell wall components of the target bacterium, and configuration of the substrate (peptidoglycan) The range of bacteria...

Ngày tải lên: 30/03/2014, 02:20

... TRH-DE was solubilized and puri ed by following exactly the procedure elaborated for the puri cation of the particulate TRH-DE from rat and pig brain [28] For the puri cation of the serum TRHDE fractionation ... the concentration of albumin and the activity of the TRH-DE in the culture medium increased in a correlative manner (Fig 2) Puri cation of the TRH-DEs from serum and liver The membrane-bound ... followed by Western blotting The proteins were again identi ed by use of the monoclonal antibody 41H2 Western blot analysis To verify the hypothesis that the brain TRH-DE, the serum TRH-DE and the...

Ngày tải lên: 31/03/2014, 21:21

... 5¢ adapters of the double stranded cDNA PCR conditions and followed procedures were identical to those used during sequencing of the 3¢ end cDNA-sequences obtained were submitted to the GenBank ... Fractionation and puri cation of the enzymes stored in the latex of Carica papaya J Chrom B 790, 229–238 Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage ... for the first time, we are not aware of the frequency of tapping in other studies These and probably several other, as yet unknown, factors affecting the activity and concentration of latex proteins...

Ngày tải lên: 07/03/2014, 11:20

... post-translational modifications of the subunit proteins Cloning and sequencing of turtle GST subunits would confirm their size, identify potential sites of post-translational modi cation and establish their place ... used and pH was confirmed immediately prior to and following the assay; the pH values shown are the average of these two values Peak (d, lines) and Peak (s, dotted lines) GST had pH optima of ... filtration and ion exchange The puri cation scheme developed for turtle GSTs resulted in final specific activities of 7.31 and 55.4 UÆmg)1 protein for Peaks and GST from aerobic control liver and 2.80 and...

Ngày tải lên: 23/03/2014, 15:20

... puri ed and characterized in this study demonstrated many properties appropriate to a variety of industrial applications The enzyme is stable for extended periods of time in the presence of calcium ... from the slope of the absorbance curve, using software installed by the manufacturer (Beckman Coulter) The extinction coefficient under the conditions described was 17454 LÆmolÆcm)1 [6] One unit of ... measured using standard conditions described in the text Results are expressed as the percentage of activity with no addition of solvent Standard error for all experiments is less than 5% of the...

Ngày tải lên: 23/03/2014, 21:20

... with the codon for Pro2 of the receptor cDNA The codon for the last used amino acid of the receptor (Ser412 in case of the full-length receptor and Ala316 for the truncated receptor) was followed ... this puri cation, 7% of speci®c ligand binding sites loaded onto the XAC-agarose were detected in the ¯ow through fraction The puri cation was carried out as outlined in the Materials and methods ... concentration of 0.3 mM and the temperature was reduced to 22 °C Cells were harvested 22± 28 h later, frozen in liquid nitrogen, and stored at )70 °C Synthesis of XAC-agarose gel The ligand af®nity...

Ngày tải lên: 24/03/2014, 00:21

... acid The substrate analogues, presumed products, and phosphate ion were eluted at the beginning of the run and collected by hand The mixture was dried by evaporation under vacuum and redissolved ... Analyzer, Applied Biosystems, CA, USA) to confirm that no unwanted mutation had been introduced The procedures for production and puri cation of the mutant protein were the same as those of the wild-type ... were gel-puri ed and assembled in a second round of PCR using oligonucleotides 40 and 41 as primers and cotemplates The PCR product was puri ed, restricted with SpeI and BamHI, and ligated with SpeI/BamHI-restricted...

Ngày tải lên: 30/03/2014, 15:20

... Kunitz-modules of the human and fugu WFIKKN proteins (wfikkn_hu_2; wfikkn_fugu_2) with the second Kunitz-domains of the human and the two fugu WFIKKNRP proteins (wfikknrp_hu_2; wfikknrp1_fugu_2; wfikknrp2_fugu_2), ... sequencing system showed that the elastase cleavage occurred at the boundary of the b-galactosidase region of the b-gal fusion protein The amino acid sequence of the resulting puri ed protein was RTDACVLPAVQGPCRGWEPRWAYS ... [8–10]) Thermal unfolding of the protein was monitored at 203 nm at a heating rate of 60 °CÆh)1 Effect of the recombinant protein on the activity of proteases The activity of the proteases on synthetic...

Ngày tải lên: 31/03/2014, 01:20

... competitors completely abolished the binding of the labeled DR4 No reduction of binding was observed when unlabelled nonspecific oligonucleotides were used (Fig 9) The order of preference for SmNR1 ... acids at the 5¢ end of the DBD, the DBD and the 40 amino acids at the 3¢ end of the DBD of SmNR1 (Ile247-Ser372) and SmRXR1 (Glu251-Asn376) were tested in vitro Lanes 1, 5, 9, 13, 17, 21, 25 and 29, ... most NRs, and the length of the hinge in SmNR1 thus can be estimated to be 140 amino acids The role of the large hinge in schistosome NRs remains unknown The degree of conservation of the LBD...

Ngày tải lên: 07/03/2014, 11:20

... and analysis of homologous sequences supported the classi cation of Bcp2 in the family of peroxiredoxin in the 1-Cys Prx class and Bcp2 could be considered the first ancient Prx developed in the ... removal of the N-terminal methionine To assess the quaternary structure of the enzyme, analytical gel filtration on PC 75 and Biosep-SEC-4000 of puri ed rBcp2 were performed The protein was eluted ... periods of time (Fig 3) As shown, the addition of stressors to the cultures inhibits growth without killing the cells The hybridizing band in the northern analysis showed the expected size of about...

Ngày tải lên: 07/03/2014, 12:20

... major fraction of the proteins in the cell culture medium, and were puried by a combination of ion exchange and gel ltration chromatography Analysis by reducing SDS PAGE showed that the apparent ... described previously [55] Expression and purication of the recombinant N-terminal region of FXII and domain D5 of HK Expression vectors for the recombinant N-terminal region of FXII and domain D5 of ... designated triafestin Therefore, the gene products of clones Ti369 and Ti263 were designated triafestin-1 and triafestin-2, respectively To study the phylogeny of these triafestins, the amino...

Ngày tải lên: 16/03/2014, 05:20

... (subset figure in a box) Recognition of proteins containing CP2-binding motifs and veri cation of the CP2-binding ability of REST and YY1 that contain the HXPR motif As stated above, none of the 12-mer ... between HA–CP2 and the puri ed peptides Whole-cell extracts from 293T cells expressing HA–CP2 were mixed with each of the puri ed GST-tagged peptides and the complex formation was analyzed by GST ... bacteria, the phage resuspension in TBS was heattreated at 70 °C for 15 The collected phage were then subjected to five more rounds of ampli cation as above The phage titer of every round was determined...

Ngày tải lên: 16/03/2014, 18:20

... equilibration of ligand ⁄ receptor at the needle tip Heats of dilution for the ligands were determined in control experiments, and these were subtracted from the integrated data before curve fitting ... 1-nm bandwidth, 1-nm resolution, 20-nmÆmin)1 scan speed and a response time of s, averaging five scans Gel filtration Two-hundred and fifty microlitres of the wild-type enzyme and 250 lL of the K142A ... composed of residues from both subunits Lys142 from one subunit contributes to the active site of the other subunit, and we therefore investigated whether its mutation (K142A) had affected either the...

Ngày tải lên: 22/03/2014, 21:21

... within the glycine residue, respectively with relay between the carbonyl carbon and the CH2 protons of the glycine moiety, thus confirming the identi cation of glycine NMR provided evidence for the ... GlcNAc residue are indicated in the inset figures also performed on two other meningococcal strains of clinical origin and the compositions of the glycoforms observed are listed in Table In each case ... LPS The identi cation and localization of the amino acid glycine substituting the HepII molecule has revealed the potential for further complexity at this inner core residue From a survey of the...

Ngày tải lên: 23/03/2014, 21:21

... completely inhibited collageninduced platelet aggregation and also attenuated the change of platelet shape Identi cation of the target receptor for triplatin-1 and -2 Next, we attempted to identify triplatin-1 ... number of inhibitors of this type have been characterized LAPP [19–21] and calin [22] were identi ed from Haementeria of cinalis and Hirudo medicinalis, respectively They prevent both the binding ... molecular masses of puri ed recombinant triplatin-1 and -2 were approximately 17 kDa on SDS ⁄ PAGE, which agreed with their predicted molecular masses of 17.9 and 17.1 kDa in secreted form To identify...

Ngày tải lên: 30/03/2014, 10:20

... molecules identi ed from the salivary gland are secreted proteins Predicted secreted proteins were classi ed into several protein families based on amino-acid sequence similarities (data not shown) The ... and (Fig 1A,B) They exhibited no sequence similarities to any other previously identi ed proteins and shared 79% sequence identity (Fig 1C) The cDNA of madanin and contained 240 and 243 bp ORFs ... thrombin Madanin and also inhibited the activation of thrombin-catalyzed cofactors V and VIII as well as the conversion of fibrinogen into fibrin Furthermore, they inhibited thrombin-induced platelet...

Ngày tải lên: 31/03/2014, 01:20