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... England Biolabs) and observed on 2.0 % agarose gel (Promega) Digested products for T1762A1764 dual mutations would yield 197 bp and 110 bp while A1762G1764 wild-type would be undigested and remained ... EcoNI (New England Biolabs) and observed on 2.0 % agarose gel (Promega) T1858 variant would produce 20 bp and 190 bp restriction products while C1858 variant would be undigested and remained at ... in Japan and Ba which is found in some other Asian countries such as China, Hong Kong, Taiwan, Thailand and Vietnam The Ba strain has recombination with genotype C in the precore region and the...

Ngày tải lên: 02/11/2012, 10:14

... lowered monocyte surface expression of HLA-DR observed in these septic patients [14] Lowered HLA-DR expression has been associated with impaired monocyte function and restoring expression to normal ... days 3, 7, and 14 for PCR studies and days 0, 1, and for measurement of soluble HLA-DR Blood (7.5 ml) was collected in preservative free heparin (final concentration of 100 iu/ml) and cells were ... cultured with GM-CSF at concentrations of 1ng/ml and 10ng/ml At time points 4, 6, and hours, cells were evaluated for surface expression of HLA-DR and HLA-DR mRNA production Control samples were...

Ngày tải lên: 03/11/2012, 09:57

... Jun 21:22 really_big.log The following command is one, cp /dev/null really_big.log is another, and echo > really_big.log is yet another All of these commands overwrite a large file with nothing ... code first defines the file in question and then checks whether it exists If it does exist, execution continues CHAPTER 40 ■ REMOVING LARGE FILES AND LOG ROLLING #!/bin/sh LOGFILE=./my.log if ... main addition to this idea here is the cp -p command cp -p $LOGFILE ${LOGFILE}.0 > $LOGFILE fi This command preserves the original file’s ownership and time stamp for the new copy Thus the new copy...

Ngày tải lên: 05/10/2013, 08:51

... decreasing protein production and gene expression for IL-1b and TNF-a [11] To address these paradoxes of both pro- and anti-inflammatory effects, the secretion of IL-1b and TNF-a from MSCs under ischaemic ... pro-IL-1b and TNF-a expression in MSCs stimulated by hypoxia ⁄ SD for the indicated number of hours (B) Representative western blots for pro-IL-1b and TNF-a expression in MSCs in the presence and absence ... 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...

Ngày tải lên: 18/02/2014, 04:20

... supplied with the enzyme, and contained Aba, Abb and Abc at 40 nm each, and the start and stop primers Abstart and Abstop at 600 nm each, and 200 lm each of dATP, dCTP, dGTP and dTTP The product was ... Ab(1–40) and Ab(M1–40) are at least 97% pure In the lanes containing Ab(1–42) and Ab(M1– 42), there were prominent bands at approximately kDa and faint bands at approximately 14 kDa The band at ... by side with Ab(1–40) and Ab(1–42) synthesized using standard Fmoc chemistry and isolated by SEC as described above Thioflavin T binds to Ab fibrils and protofibrils [30], and has been extensively...

Ngày tải lên: 18/02/2014, 13:20

... constructs of Caveolin-1 (shRNA1 and shRNA2) and three different scrambled shRNAs (control 1–3), cells were harvested, and the expression levels of Caveolin-1 and of P2X7 receptor were analyzed ... nonspecific band However, immunoreactivity to both the 80 kDa and 60 kDa bands was abolished following preincubation with the antibody control peptide (Fig 1D), implying that the 60 kDa band shared ... Analysis of P2X7 receptor expression in alveolar epithelial lung cell lines P2X7 receptor expression was measured in cell lysates of A549, E10, R3 ⁄ and L2 cells by SDS ⁄ PAGE and western blot analysis...

Ngày tải lên: 19/02/2014, 00:20

... curves and data were normalized to the copy number of 18S RNA Expression of ACMSD isoforms in E coli pGEM-ACMSD I and pGEM-ACMSD II were digested with XhoI and Bpu1102I and cloned into pET-15b and ... I at the N-terminus (Fig 2B) Expression of ACMSD variants in Escherichia coli and Pichia pastoris ACMSD I and II cDNAs were subcloned into pET15b and pET32b E coli expression vectors, providing ... swiss-pdbviewer and visual molecular dynamics [49] Acknowledgements We thank G Magni and S Ruggieri for helpful comments and suggestions and T Begley (Department of Chemistry and Chemical Biology,...

Ngày tải lên: 19/02/2014, 02:20

... 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 ... a template, and the PCR products were subjected to 2% agarose-gel electrophoresis, and stained with ethidium bromide (C) Expression of Noxo1b and Noxo1c in human adult tissues The expression levels ... Noxo1b, Noxo1c, Nox1, and Noxa1 in various human cell lines: androgen-independent prostate cancer cells (LNCaP), androgen-independent prostate cancer cells (PC3 and DU145), and testicular germ...

Ngày tải lên: 19/02/2014, 06:20

... parasites after days and 2, 4, and 24 weeks after transfection Lanes 1, 3, 5, 7, 9, 11 and 13 show the PCR products obtained using the primer pair T7 and RIB Lanes 2, 4, 6, 8, 10, 12 and 14 show the ... discussions and Edith Trejo and Carlos Zadikian for technical assistance We are indebted to Drs J J Cazzulo and C Labriola for their generous gifts of a cruzipaincontaining plasmid and primers A and ... after electroporation Hybridization bands with ADC- and rRNA-specific probes are shown in (A) and (B), respectively (C) Relative intensities of ADC mRNA bands normalized to the rRNA loading controls...

Ngày tải lên: 19/02/2014, 07:20

... specific for human Ras-GRF1 Primer pairs: lane 2, ON359 and ON360; lane 3, ON357 and ON358; and lane 4, ON361 and ON360 The PCR products and a DNA size marker, lane 1, were separated on agarose ... increase and show Ca2+ dependence of serotonininduced ERK1 ⁄ phosphorylation and a mandatory role of the Ca2+ ⁄ CaM-binding IQ domain in RasGRF1-stimulated ERK1 ⁄ phosphorylation Thus, both cAMP and ... that the treatment of NIH3T3 and COS-7 cells with carbachol [9] and lysophosphatidic acid [11], activating both Gq- and Gi-coupled receptors, induces the activation and phosphorylation of Ras-GRF1...

Ngày tải lên: 19/02/2014, 18:20

... x-5a and x-5b, of 1.4 and 1.2 kb, respectively (Fig 1) Both genes were cloned into Escherichia coli XL-10 Gold and the nucleotide sequence was determined The x-5a gene consists of 1413 bp and ... Cloning and expression of wheat x-5 gliadin at positions 288 and 1170 The nucleotide sequences obtained from this study have been deposited in the DDBJ database under accession numbers AB181300 and ... H Matsuo et al Cloning and expression of wheat x-5 gliadin Fig Western blot analysis of native and recombinant x-5 gliadin with IgE antibodies from patients with WDEIA and healthy controls One...

Ngày tải lên: 20/02/2014, 01:20

... four-band pattern at SDS/PAGE (Fig 1) At comparison with insect cell produced recombinant cystatin F (Fig 1, lanes and 2), which appears as one mono- (band B) and one di-glycosylated (band A) ... nuclei and cell debris), a heavy and a light fraction containing mitochondria and various cell organelles, and a supernatant containing the smallest cellular vesicles Cystatin F, cystatin C and ... reported The expression of the cystatin F (CST7) and C (CST3) genes differ considerably, both with respect to the overall levels and with respect to tissue/cell-specific expression pattern CST3 expression...

Ngày tải lên: 21/02/2014, 01:21

... activation and NAMI-A- induced down regulation of c-myc gene expression NAMI-A inhibits c-myc gene transcription and protein expression To establish whether the decrease in c-myc mRNA expression ... c-myc gene expression with superimposable magnitudes and kinetics profiles Figure 5C shows the dose–response effect of NAMI-A on ECV304 c-myc mRNA expression As reported for DNA synthesis and ERK1/2 ... immunoreactivity of ERK1/2 and phospho ERK1/2 (A,B), and MEK1/2 and phospho MEK1/2 (C,D) The lower part of each panel reports the quantitative analysis of phospho ERK1/2 (A,B), and phospho MEK1/2 (C,D)...

Ngày tải lên: 21/02/2014, 01:21

... mutagenesis MATERIALS AND METHODS Expression vector The FLS cDNA from satsuma mandarin fruits, C unshiu [30], was excised with EcoRI and XhoI from the Bluescript vector (Stratagene) and subcloned in ... this enzyme has not been functionally proved and the lability of substrate and product rule out any cocrystallization The setting of a helices and b-strands causes very similar CD spectra for this ... deoxycholate [40] and using bovine serum albumin as a standard Flavonol synthase (Eur J Biochem 269) 4137 and Matthiola incana), but functionality was only verified in case of Petunia and Arabidopsis...

Ngày tải lên: 21/02/2014, 03:20

... heterologous expression system for hevamine in Escherichia coli, and used X-ray analysis and enzyme kinetic experiments to gain detailed insight in the role of these residues in catalysis MATERIALS AND ... Sense strand 5¢-GATGGTATTGATTTTGCCATAGAGCATGGTTCA-3¢ Anti-sense strand 5¢-TGAACCATGCTCTATGGCAAAATCAATACCATC-3¢ Sense strand 5¢-TTGGATGGTATTGATTTTAACATAGAGCATGGTTCAACC-3¢ Anti-sense strand 5¢-GGTTGAACCATGCTCTATGTTAAAATCAATACCATCCAA-3¢ ... strand 5¢-GGTATTGATTTTGACATAGCGCTATGTCAAAATCAATACC-3¢ Anti-sense strand 5¢-GTACAGGGTTGAACCATGCGCTATGTCAAAATCAATACC-3¢ Sense strand 5¢-GATGGTATTGATTTTGCCATAGCGCATGGTTCAACCCTG-3¢ Anti-sense strand...

Ngày tải lên: 22/02/2014, 04:20

... using various substrates and compared with those of hPDI The recombinant cPDI and hPDI shared similar reductase, oxidase and isomerase activities PDI exhibits both chaperone and antichaperone activities ... Cl, pH 7.5, mM EDTA) The filled and open circles denote native and linear tx3a, respectively (B) Refolding carried out in buffer B (buffer A plus mM GSH and mM GSSG) and in buffer C (buffer B plus ... into an 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

... of the WISP1v proteins in HCS-2 ⁄ cells and their overexpression (A) Molecular constructs for the overexpression of WISP1, WISP1v and WISP1vx A mammalian expression vector, pFLAG-CMV was employed ... outcome of the constitutive expression of the full-length and the differentiation stage-dependent expression of WISP1v in vitro Effect of overexpression of CCN4/WISP1 and its variants on the phenotypes ... enhanced expression evaluated by real-time RT-PCR; ±, modest expression evaluated by real-time RT-PCR; –, expression undetectable ticular, expression of WISP1v among normal chondrocytes and osteoblastic...

Ngày tải lên: 07/03/2014, 10:20

... those concepts and aesthetic concepts XIII THE SO-CALLED PHYSICALLY BEAUTIFUL IN NATURE AND IN ART Aesthetic activity and physical concepts Expression in the aesthetic sense, and expression in ... for action and his small amount of theoretical clarity as regards the situation and the means to be employed And if he develop a taste for contemplation and discovery, and leave willing and acting, ... sciences, and their limits–The phenomenon and the noumenon IV HISTORICISM AND INTELLECTUALISM IN AESTHETIC Critique of the verisimilar and of naturalism–Critique of ideas in art, of art as thesis, and...

Ngày tải lên: 07/03/2014, 12:20

... [20,22] and Bacillus subtilis lipase A [23] Results and Discussion Cloning and expression of Rv1399c The Rv1399c gene product encodes a 318 amino acid protein with a molecular mass of  31.7 kDa and ... 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 on ... 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...

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 ... SMEZ1 and 2), three have one Cys (SEI, SEK and SPEC), two have three Cys (SEG and SPEA) and only SSA has more than that, five Cys As discussed later, the fact that SSA has two Cys residues (Cys26 and ... NaCl/Pi and concentrated to mgÆmL)1 hVb2.1hCb2 and hVb1hCb2 were also produced as inclusion bodies and refolded at pH 8.5 Purification steps included gel filtration on a Superdex 200 FPLC column and...

Ngày tải lên: 07/03/2014, 16:20