expression and purification of rhupon1 from e coli

... of cHSPA6 overexpression in E Coli Effect of temperature on the overexpression of cHSPA6 Figure showed that 37 °C was found to be the optimum temperature and further optimization was performed ... cultures is shown in Table These results showed that the optimal induction at the mid exponential phase produced high levels of soluble proteins with high cell density Figure Effect of pre-induced ... temperature 2.2.2 Culture media optimization LB media showed relatively higher growth rate under induced culture conditions resulting in higher volumetric yield Figure Effect of media on the...

Ngày tải lên: 24/10/2016, 14:24

... the presence of impurities could not be excluded Therefore, reproducible and reliable methods for separation and purification are required for the accurate characterization of these enzymes The ... dehydrogenase in presence of FMN and remove electrons from NADH to catalyze the reaction (Trudgill 1986) Low activities of purified oxygenating component were observed earlier as well and were explained ... cultivation Gene expression analysis Gene expression analysis was performed with crude cell extract Samples standardized to cell amount were taken during cultivation Cells were harvested by centrifugation...

Ngày tải lên: 21/06/2014, 02:20

... antibodies directed to non-native epitopes represented in the bacterial-expressed antigen None of the recombinant LASV proteins were specifically detected by sera from Lassa fever naïve donors ... gene sequences were restricted and cloned in-frame at the 3' end of the malE gene, beyond the cleavage site for Factor Xa (IQGR) The LASV GP1 gene sequence comprised a.a 59–259 in the native GPC, ... Nurse, Cancer Research UK Your research papers will be: available free of charge to the entire biomedical community peer reviewed and published immediately upon acceptance cited in PubMed and...

Ngày tải lên: 20/06/2014, 01:20

... the role of cysteine protease in this, we used the cell permeable cysteine protease inhibitor E- 64d Following infection of insect cells with vORF1, the cells were treated with E- 64d and the cell ... been demonstrated We reasoned this may be due to improper folding or low expression levels of the polyprotein in subgenomic expression systems attempted so far We show here expression of the ... Nine tryptic peptides were selected from the mass spectrum (Fig 5A) and compared for their experimentally obtained and predicted masses (Fig 5B) These predicted sequences matched the N- terminal...

Ngày tải lên: 20/06/2014, 01:20

... the chimeric protein retained heightened immunogenicity based on the adjuvant properties of the genetically linked RTB Indeed, the presence of RTB markedly enhanced immune responses in mice to p24 ... Nurse, Cancer Research UK Your research papers will be: available free of charge to the entire biomedical community peer reviewed and published immediately upon acceptance cited in PubMed and ... samples were diluted at 1/200, and levels of IgG anti-p24 antibodies were determined by ELISA All sera samples were tested in triplicate and standard deviations are included strong immune response...

Ngày tải lên: 12/08/2014, 04:21

... residues generates a more sensitive enzyme, whereas the absence of any glutamine residue in the active site renders the enzyme more sensitive) The reason for this is unclear Azide, like hydrogen peroxide, ... FEBS 2002 5146 T Hunter et al (Eur J Biochem 269) levels of hydrogen peroxide has therefore been used as an indicative marker for the presence of iron at the active site and, indeed, is frequently ... ion from which the SOD derives, and very little of the contrary metal is present (Table 2) This result demonstrates the selectivity of each of the wild-type enzymes for its respective Table Metal...

Ngày tải lên: 21/02/2014, 01:21

... 2Fo-Fc electron density maps were calculated and inspected with O [30] to check the agreement of the model with the data RESULTS Expression of hevamine in E coli Initially, we tried to use an expression ... between colloidal chitin and the enzyme influences the active site properties The cause of these differences is not known Km and kcat measurements of hevamine and mutants Comparison of the steady-state ... METHODS Heterologous expression of hevamine in E coli For the heterologous expression of hevamine in E coli, the T7 based expression vector pGELAF+ was used [18] A construct, named pHEV, was made,...

Ngày tải lên: 22/02/2014, 04:20

... data are the average of three independent experiments For the rate of increase of the native form, the original data were fitted by Y(t) ¼ Ymax(1 ) e kt) · 100%, where Y is the percentage of the native ... three independent experiments For the rate of decrease of the linear form, the original data were fitted by Y(t) ¼ e kt · 100%, where Y is the percentage of the linear form, and t is the refolding ... the presence or absence of 200 gÆL)1 Ficoll 70 The amounts of nTx3.1 and sTx3.1 were calculated from their elution peak areas The data are the average of three independent experiments For the...

Ngày tải lên: 07/03/2014, 05:20

... Substrate concentrations (indicated in parentheses) are expressed in mM; ND, non determined Acetylcholinesterase and pig liver esterase values are taken from Table of reference [7] Substrate (concentration) ... we have established that Rv1399c is a NLH, consistent with the sequence homology detected between Rv1399c and other serine carboxyl ester Table Flowsheet of the Rv1399c procedure purification and ... pH and temperature dependence experiments rather than more volatile vinyl esters The pH stability profile of Rv1399c was obtained after enzyme incubation for h using 100 lL of buffers at different...

Ngày tải lên: 07/03/2014, 16:20

... with the selectivity (K2/K1) or the difference between K2 and K1, and the reduction in band width Within the eight tested solvents systems, the differences between K2 and K3, and between K4 and ... Petroleum ether/acetonitrile/ acetone (2:1:1) Petroleum ether/acetonitrile/ acetone (5:3:2) Petroleum ether/acetonitrile/ acetone (4:3:1) Petroleum ether/acetonitrile/ acetone (7:6:1) n-Heptane/acetonitrile/ethyl ... (Table 1) These two solvent systems (petroleum ether/ acetonitrile/acetone 4:3:1 and 2:1:1 v/v/v) were selected for further study The settling times for solvent systems of petroleum ether/acetonitrile/acetone...

Ngày tải lên: 07/03/2014, 21:20

... (d.p.m.) Cell-free homogenate In the absence of WBP E coli BL21(DE3)/pET32b E coli BL21(DE3)/pETHRT1 E coli BL21(DE3)/pETHRT2 In the presence of WBP WBP E coli BL21(DE3)/pET32b + WBP E coli BL21(DE3)/pETHRT1 ... homogenates of E coli BL21(DE3)/pETHRT1 and E coli BL21(DE3)/pETHRT2 were examined for in vitro rubber transferase activity Rubber materials were extracted with a solvent mixture of toluene and hexane ... reactions were performed using Thermo Sequenase Cycle Sequencing kit (Amersham Pharmacia Biotech) with fluorescent labeled primers Nucleotide sequences were determined by the dideoxy chain termination...

Ngày tải lên: 07/03/2014, 21:20

... paper were displayed purple in color To determine the effects of pH, temperature and inhibitors, l-aspartate and a-ketoglutarate were used as amino donor and acceptor, respectively, and the reactions ... biosynthesis of the lipopeptides [26] We presume that the aat gene in this region can regulate the biosynthesis of the lipopeptides The experiments performed in the present study showed that ... a-COO) of the substrate plays a key role in the activity of the enzyme [19,20] The functions of the above-mentioned conserved active residues were all identified by using the AAT from E coli as the...

Ngày tải lên: 14/03/2014, 23:20

... four two-residue insertions, one three-residue insertion, one six-residue insertion, and one 19-residue insertion; there are two oneresidue deletions From this alignment, seven conserved regions ... functional homologies of this enzyme between invertebrates and vertebrates Whereas the sequences of the N-terminus and C-terminus of the Conus carboxylase are quite divergent, we found significant ... N-terminal and C-terminal sequences Alignment of the Conus carboxylase sequence with the primary structure of the vertebrate carboxylase homologs indicates the presence of one one-residue insertion,...

Ngày tải lên: 17/03/2014, 10:20

... have the same evolutionary origin and have divergently evolved to exhibit different peptidase specificities Site-directed mutagenesis of the putative active-site residues to other proteinogenic ... active-site residues of PepV and CPG2 The active-site residues are indicated by gray, yellow and cyan for PepD, PepV and CPG2, respectively The active-site residues are almost equivalent The zinc-binding ... enzymes have been studied extensively, the functional residues of PepD-related enzymes are poorly understood To determine the importance of PepD in affecting biofilm formation and serving as a potential...

Ngày tải lên: 23/03/2014, 06:20

... innate immune system, would be of considerable interest Before the present study was started, only vertebrate 2-5A synthetases had been expressed in heterologous systems for use in detailed studies ... the presence of the nuclease Inspection of UV-spectra of the nuclease-treated and untreated preparations revealed that both of them were contaminated with nucleic acids Calculation of RNA content ... d, recombinant protein preparation, where the number of nucleotides was decreased by nuclease treatment The number of nucleotides per protein monomer was estimated as described in Experimental...

Ngày tải lên: 23/03/2014, 09:20

... CMCase/xyloglucanase) was added to a mixture of EXOIII and EndoI The amount of cellobiose released was twice as high as the amounts released by EXOIII and EndoI alone; however, even the most effective ... cuts cleanly and Xeg74 does not, as both genes are cloned in the same way except for the presence of the linker and the family-2 CBM at the C terminus of XEG74 The molecular masses of Cel9A-68 ... aculeatus enzyme (FIII avicelase), which produces cellotriose and cellobiitol from reduced cellopentaose, clearly showing that it releases cellobiose from the reducing end [30] However, the enzyme was...

Ngày tải lên: 23/03/2014, 21:20

... be partially achieved by expression of the ECDs of the AChR subunits in heterologous expression systems Several studies have been carried out on the expression of the muscle-type a subunit ECD ... [18] Based on this experience, we embarked on the expression of three of the four non-a ECDs, namely b, c and e The expression of the d subunit ECD, which is currently under progress, presents major ... significant effect on expression (less than 10%) The presence of the FLAG tag increased expression of c1–218 by almost 20%, but did not improve expression of c1–219 The presence of the HIS tag alone reduced...

Ngày tải lên: 30/03/2014, 10:20

... site (Fig 1) To investigate further whether there are two related STAT5 genes in the pufferfish genome, we used a pair of degenerate primers, which were derived from two highly conserved regions ... The sequences of the upper strand of the normal and the corresponding mutant oligonucleotides are listed in Table The probes were prepared by annealing the upper and lower strands of oligonucleotides, ... All protein sequences were aligned pairwise using the Geneworks nucleic acid and protein sequence analysis software 2.5 from Intelligenetics, Inc The numbers represent percent amino acid identity...

Ngày tải lên: 31/03/2014, 07:20

... exonuclease III by the reported method [24] to generate a set of nested deletions from each end of the inserts Appropriately deleted derivatives were sequenced by the dideoxy chain termination method ... nitroethane) and was finally maintained in a 0.6-mL volume Nitrite released from the reaction was determined by the method of Little [28] One unit of NaoA is defined as the amount of enzyme required ... nitroalkane-oxidizing enzyme from Streptomyces; partial purification of the related enzyme from Streptomyces has been reported [16] The deduced aminoacid sequence of NaoA from S ansochromogenes has high identity...

Ngày tải lên: 31/03/2014, 08:20

... < 0.001) Results are representative of three separate experiments LPS After one day of treatment, luciferase levels indicated an induction of the presumptive serine racemase promoter by both ... microglia-conditioned medium that had been pre-treated with DAAOx Values represent the mean ± SEM of triplicate determinations, and the results are representative of three experiments (*p < 0.01 versus "no ... blocked the effects on neuronal [Ca2+]i Aβ treatment elevated the steady-state levels of serine racemase mRNA and protein, suggesting that increased synthesis may be involved in the release of...

Ngày tải lên: 19/06/2014, 22:20