expression and purification of idps

... the classification of LASV as a National Institutes of Allergy and Infectious Diseases (NIAID) Category A pathogen and biosafety level-4 (BSL4) agent Expression and purification of E coli-generated ... 2(DE3) cells transformed with construct pMAL-c2x:NP Expression and purification of LASV NP from E coli Rosetta Expression and purification of LASV NP from E coli Rosetta 2(DE3) cells transformed ... transformed with construct from E coli Rosetta gami cellsand purification of LASV GP2pMAL-c2x:GP2 Expression Expression and purification of LASV GP2 from E coli Rosetta gami cells transformed with...

Ngày tải lên: 20/06/2014, 01:20

... protein One unit is defined as the amount of enzyme that catalyzes the oxidation of μmol of substrate per minute Results Cloning, expression and purification of 2, 5-diketocamphane 1,2-monooxygenase ... simplified purification via affinity chromatography and characterization of the enzyme Material and methods Enzymes, chemicals and media Pfu+-polymerase was obtained from Roboclon (Berlin, Germany) and ... was introduced using the sites of restriction endonucleases NdeI and XhoI for cloning with the BCA-kit and a standard curve of BSA in the same buffer in a range of 2-0.005 mg/mL was used Samples...

Ngày tải lên: 21/06/2014, 02:20

... shown in Fig 7b, the level of cHSPA6 expression reached at its maximal level within 1h of induction and incubation at 37 oC The level of cHSPA6 remained unchanged up to 24h of post-induction incubation ... allows very sensitive detection (1–10 ng of protein per band) with negligible background staining 2.1 Results Expression of recombinant cHSPA6 The result express of recombinant cHSPA6 vector which ... were induced with varying concentrations of IPTG (0, 10, 25, 50, 100, 250, 500 and 1000 μM) and further expressions were made for h at 37 °C An equal amount of soluble protein extract was analyzed...

Ngày tải lên: 24/10/2016, 14:24

... in order to secrete IL-1b and TNF-a Hypoxia ⁄ SD induces the transcription and secretion of IL-10 Because of the lack of secretion of the inflammatory cytokines IL-1b and TNF-a from hypoxia ⁄ SD-stimulated ... presence and absence of IL-10 *P < 0.05 and **P < 0.01 versus Cont group (D) Representative western blots for collagen I and III in the presence and absence of 0.1 lM angiotensin II and IL-10 ... effects of MSCs in vitro mediator of the cells’ paracrine anti-fibrotic effects These findings help to improve our understanding of the cellular and molecular basis of MSCs’ anti-inflammatory and paracrine...

Ngày tải lên: 18/02/2014, 04:20

... compilation ª 2006 FEBS 3673 Expression and function of Noxo1c R Takeya et al Expression of Noxo1b and Noxo1c in various human tissues The expression patterns of the Noxo1b and Noxo1c messengers were ... compilation ª 2006 FEBS 3665 Expression and function of Noxo1c R Takeya et al Expression of Noxo1b and Noxo1c in various human tissues To compare the distribution pattern of Noxo1b and Noxo1c, we designed ... Sciences, Turku, Finland) Estimation of expression of cytosolic regulatory proteins Total cell lysates of transfected CHO cells were used to estimate expression of Noxo1b, Noxo1c, and Noxo1a The lysates...

Ngày tải lên: 19/02/2014, 06:20

... properties The cause of these differences is not known Km and kcat measurements of hevamine and mutants Comparison of the steady-state kinetic parameters of hevamine and the Tyr183Phe and Asp125Asn ... in the role of these residues in catalysis MATERIALS AND METHODS Heterologous expression of hevamine in E coli For the heterologous expression of hevamine in E coli, the T7 based expression vector ... Total number of observations Number of unique reflections Completeness (%)

Ngày tải lên: 22/02/2014, 04:20

... integrated elution peak areas, and the PDI’s oxidase and isomerase activities were expressed as the initial rate of decrease of linear tx3a and the initial rate of increase of nTx3.1, respectively To ... 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 time For the rate of increase of the ... expression vector, pET24a, which contains an N-terminal His6 tag Expression and purification of cPDI and hPDI The expression plasmid of cPDI was transformed into BL21 (DE3) cells The transformed...

Ngày tải lên: 07/03/2014, 05:20

... 205 and 215 nm, indicating that Rv1399c is composed of 22% of a-helices, 25% of b-strand and 39% of random coil The catalytic triad is located at the bottom of a small ˚ ˚ pocket, 20 A length and ... sheet is composed of parallel b strands associated to an antiparallel strand (b2) and is surrounded by helices (a1, a2, a3, a7 and a8) The second domain consists of helices a4, a5 and a6 all clustered ... in LB medium after h of IPTG induction (Fig 1) Purification and refolding of Rv1399c Given the large quantity of insoluble material obtained [up to 160 mg of protein from L of cell culture (Table...

Ngày tải lên: 07/03/2014, 16:20

... hVb2.1hCb2 and hVb1hCb2 (genes kindly provided by U Utz and R P Sekaly, University of Montreal, Canada), were cloned between the NdeI and EcoRI restriction sites of the pET17b expression vector and ... pathologies and because of the potential use of the toxins as biological weapons Alteration of their MHC and TCR binding capacity by site directed mutagenesis could be useful in the development of vaccines ... 30 and centrifuged for 10 at 12 000 g The supernatant was saved on ice and the pellet was resuspended in 50 mL of a : dilution of Tes and centrifuged as before Both supernatants were mixed and...

Ngày tải lên: 07/03/2014, 16:20

... total of mg of curdione (1), mg of curcumol (2), 10 mg of germacrone (3), 18 mg of curzerene (4), mg of 1,8-cineole (5) and 17 mg of b-elemene (6) from the essential oil (300 mg) in 2.4 mL of the ... the differences between K2 and K3, and between K4 and K5 in solvent systems (petroleum ether/acetonitrile/acetone 2:1:1 and 4:3:1 v/v/v) are 0.34 and 0.24, and 0.20 and 0.20, respectively, which ... Both of the upper and the lower phases were degassed for 30 by ultrasonication before use Aliquots of 300 mg of the essential oil were diluted by 2.4 mL of the mixed solution of lower phase and...

Ngày tải lên: 07/03/2014, 21:20

... probes As shown in Fig (A1 and A2) both probes gave specific hybridization bands of kb which are consistent with the sizes of each of the cDNAs The expression patterns of HRT1 and HRT2 mRNAs among the ... HRT2 The pETHRT1 and pETHRT2 were constructed and introduced into E coli BL21(DE3) The expression of these genes were induced by addition of 0.5 mM IPTG Overexpression of HRT1 and HRT2 is shown ... yielding the expression plasmids pETHRT1 and pETHRT2 Each of the expression plasmids was used for transformation of E coli BL21(DE3), and mL of an overnight culture of the transformant in Luria–Bertani...

Ngày tải lên: 07/03/2014, 21:20

... lg of platelet protein and 0.5 lg of GST and GST±Rab proteins; (B) an [a-32P]GTP overlay [17] of a nitrocellulose blot of a gel containing 40 lg of platelet protein, 0.5 lg of GST and 0.5 lg of ... form of Rab32 (see Discussion) Fig Puri®cation and properties of wild-type and mutant Rab31 and Rab32 expressed as GST±fusion proteins Samples of platelet particulate fraction protein and of puri®ed ... cycles) and at 72 °C, for a total of 21 cycles The PCR product was puri®ed using a QIAquickTM kit and cloned into the BamHI and XhoI sites of the pGEX-4T-1 expression vector The sequence of the...

Ngày tải lên: 08/03/2014, 16:20

... subtilis and B circulans should belong to subgroup Ib Expression and purification of AATB3 and its mutants To produce recombinant AATB3 and the three mutant proteins, the aatB3 gene and its mutants ... inserted into KpnI and EcoRI sites of pET30a(+) for the expression of protein AATB3; T7 promoter-based expression vector; Kmr The aatB3 fragment was inserted into KpnI and EcoRI sites of pUC19 for ... into KpnI and EcoRI sites of pET30a(+) for the expression of protein D232N; T7 promoter-based expression vector; Kmr The fragment from pUCK270H was inserted into KpnI and EcoRI sites of pET30a(+)...

Ngày tải lên: 14/03/2014, 23:20

... surface of the PO and many nerve fiber tracts and varicosities in trunks of the PO Cloning and sequencing of PO-CHH cDNA The first PCR of PO cDNA with a combination of a set of primers LF1 and LR1 ... Expression and release of CHH in the blue crab J S Chung and N Zmora 694 Results Identification and bioactivity of CHHs from ES sinus glands and PO Two RP-HPLC peaks (1 and 2) as presented ... Chung and N Zmora Expression and release of CHH in the blue crab Fig Sequence alignments of nucleotide and deduced amino acids of full-length PO-CHH (DQ667141) and ES-CHH (AY536012) PO-CPRP and...

Ngày tải lên: 16/03/2014, 06:20

... the expression of mutant huntingtin and the exact mechanism by which mutant huntingtin alters the function of such factors This work describing the gene structure of CB1 and its pattern of expression ... levels of CB1 mRNA in the medial striatum and majority of cortical neurons remained constant in wildtype and R6/1 and R6/2 transgenic HD mice Isolated neurons that had high levels of CB1 mRNA expression ... (C and D) are shown For each series of protection reactions, 10 pg of undigested probe control (P) and a no target control that consists of 400 pg of probe combined with excess yeast RNA and...

Ngày tải lên: 16/03/2014, 18:20

... isolation of a bovine tryptase gene and cDNA encoding a new tryptase isoform (BLT), which is in turn the only one isoform expressed in bovine lung Furthermore, analysis of BLCT and BLT expression ... fraction contains 0.3 nmol of active Ó FEBS 2003 Tissue-specific expression of bovine tryptases (Eur J Biochem 270) 513 Fig Comparison of promoter sequences and 5¢-UTRs in BLT and BLCT genes Identical ... m/-z 1490.7 and 1366.2 represent the peptides that characterize BLT isoform with respect to BLCT isoform (corresponding to peptides 167–180 and 188–201, with theoretical Mrs of 1490.5 and 1365.7,...

Ngày tải lên: 17/03/2014, 09:20

... the cloning and expression of the M tuberculosis tps gene in E coli, and the production of active TPS in these cells The properties of the recombinant enzyme have been determined, and are compared ... M) of NH4HCO3 An aliquot of each fraction was removed and assayed for its radioactive content The radioactive peak was pooled and concentrated to dryness a number of times in the presence of ... identity and 6% similarity Fig shows the predicted amino acid sequence of the M tuberculosis TPS and its sequence alignment with those of homologous ORFs from several other mycobacteria and OtsA of...

Ngày tải lên: 17/03/2014, 10:20

... proprothrombin), proFIX18 (residues )18 to )1 of proFactor IX), proFIX28 (residues )18 to +10 of proFactor IX), pro-e-TxIX/12 (residues )12 to )1 of e-TxIX precursor), e-TxIX12 (residues 1–12 of e-TxIX), ... ducts, expression of the recombinant Conus carboxylase in a system free of endogenous carboxylase activity and substrates will considerably facilitate studies of the mechanistic properties of this ... FLEEL, proPT28 and proFIX28, were Table Specific carboxylase and epoxidase activity of recombinant Conus carboxylase in the absence or presence of 400 lM proPT18 ND, not determined Expression system...

Ngày tải lên: 17/03/2014, 10:20

... of the tail of injected shrimps (Fig 9D and E) Confrontation of haemocytes and bacteria The recruitment of penaeidin-positive haemocytes around the site of injection confirmed the importance of ... site of microbial injection precluded both clarification of the mechanisms of penaeidin release from haemocytes, and determination of any potential involvement of penaeidin in the elimination of ... slides per shrimp and each value represents the average of four shrimps Histograms refer to the percentage of haemocytes exhibiting more than 50 of AU (black bars) and the percentage of haemocytes...

Ngày tải lên: 18/03/2014, 01:20

... lL of mm l-carnosine (dissolved in 50 mm Tris–HCl, pH 7.4) and 20 lL of purified enzyme (0.5 mgÆmL)1,  lm) The reaction was initiated by addition of substrate and stopped by addition of 50 lL of ... mass of 53.6 kDa and an isoelectric point of pH 4.7 Production of the V alginolyticus PepD protein The V alginolyticus pepD gene was then subcloned into the expression plasmid pET28a(+) and subsequently ... citric acid (pH 4, and 6) and Tris–HCl (pH 6, 7, 7.4, 8.5, and 9.5) (Fig 4A) The pH activity of PepD showed an optimal activity in the range pH 7–7.4 and declined at more acidic and alkaline pH...

Ngày tải lên: 23/03/2014, 06:20