expression and purification of recombinant proteins for vaccine applications

... 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 ... Gami bla Figure strategy for expression of LASV proteins GP1, GP2, and NP in E coli using pMAL vectors Cloning Cloning strategy for expression of LASV proteins GP1, GP2, and NP in E coli using ... 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 LASV proteins...

Ngày tải lên: 20/06/2014, 01:20

... SiMAG/CS-NTA/Nickel for purification of two Histagged recombinant proteins, His-ProT and His-Mre11, overexpressed in Escherichia coli Results Relative expression of His-tagged recombinant proteins Recombinant proteins ... matrices is mandatory, which may result in uniform purification efficacy Performance of such commercial magnetic matrices for purification of different His-tagged proteins is, therefore, required ... insights for developing versatile and userfriendly resins suitable for purification of a vast array of recombinant His-tagged proteins Methods Instruments Magnetic separation stand and permanent...

Ngày tải lên: 11/08/2014, 00:23

... previous data for expression of the N protein of RVFV Expression of RVFV glycoproteins using the baculovirus expression system has been reported before [24,28] but functional analysis of these proteins ... could form multimeric complex and assembled into a particulate structure in the absence of genomic RNA Figure Assembly of VLPs by expression of RVFV N and Gc proteins Assembly of VLPs by expression ... expression of RVFV Gn and Gc proteins in insect cells resulted in detection of both proteins on the surface of the non-permeabilized infected cells The presence of Gn and Gc on the surface of infected...

Ngày tải lên: 20/06/2014, 01:20

... temperature for (+)-1, (-)-1, 13, 14 and 15 was 60°C for followed by a gradient of 10°C/min to 180°C maintained for Detection temperature for 16 was 120°C For 17, 240°C for followed by a gradient of 2°C/min ... purification, 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 ... shifting the culture to 20°C and 200 rpm for another 16 h of cultivation Gene expression analysis Gene expression analysis was performed with crude cell extract Samples standardized to cell amount...

Ngày tải lên: 21/06/2014, 02:20

... (residues )18 to +10 of 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), ... of either the recombinant Conus carboxylase or the recombinant bovine carboxylase, and monitored carboxylation of FLEEL or e-TxIX12 as a function of propeptide concentration The effect of proFIX18 ... 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

... procedures of the porcine 2-5A synthetase (data not shown) Figure demonstrates the results of the purification of the recombinant proteins The occurrence of dominant bands of the recombinant proteins ... sequence Expression and purification of the recombinant 2-5A synthetase from G cydonium Expression and purification of the porcine recombinant 2-5A synthetase N-terminally 6xHis-tagged construct The recombinant ... characterized by means of a recombinant protein technique Results Expression and purification of His-tagged proteins N- and C-terminally hexahistidine tagged constructs of the 2-5A synthetase...

Ngày tải lên: 23/03/2014, 09:20

... the procedure of Tunon and Johansson, 1984 This protocol allows very sensitive detection (1–10 ng of protein per band) with negligible background staining 2.1 Results Expression of recombinant ... Optimization 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 ... 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

... 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 of BAY 11-7082 or MG132 *, nonspecific band (C) ... stimulation for 12 h in the presence and absence of ATP and LPS (B) Representative western blots for TNF-a expression in MSCs stimulated by hypoxia ⁄ SD in the presence or absence of LPS for the ... 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

... conditions for the production of the I7 OR in yeast and used biochemical and immunological methods to estimate the levels of receptor expression and its cellular localization Results Yeast transformations ... Identification of ligands for olfactory receptors by functional expression of a receptor library Cell 95, 917–926 20 Hatt H, Lang K & Gisselmann G (2001) Functional expression and characterization of odorant ... 527 Expression of olfactory receptors in yeast for screening A J Minic et al and Ga15 in membrane fractions are constant regardless of the temperature and galactose induction, while the level of...

Ngày tải lên: 07/03/2014, 16:20

... 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 ... 421 of the M tuberculosis TPS Isolation and purification of recombinant TPS To maximize conditions for production of recombinant TPS, the E coli vector was incubated in 0.1 mM or mM IPTG for h ... 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...

Ngày tải lên: 17/03/2014, 10:20

... [23], and Hem O, N meningitidis A 2855 [25] Soret and Q (a and b) bands of the oxy form and the CO-bound form not show specific differences from those of mammalian or bacterial HOs The small band ... optical absorption spectra of the ferric-, ferrous-, oxy-, and CO-bound forms of heme–Syn HO-1 The Soret bands of the ferric and ferrous forms, having maxima at 402 and 427 nm, respectively, are ... water-soluble proteins and mainly function to release iron from heme of the host cells, which is necessary for the survival of bacteria and for causing diseases The crystal structure of the bacterial...

Ngày tải lên: 23/03/2014, 20:22

... were imaged for GFP-SCAP (B and E) or GM130 (C and F) Panels D and G are merged images of GFP-SCAP and the Golgi marker GM130 Scale bar, 10 lm (H) Quantification of the percentage of Golgi-localized ... of SREBP functions The most likely are, first, an increase in the gene expression of SREBPs, and second, the enhancement of SREBP processing by glutamine We therefore investigated the effect of ... effect of glutamine on the gene expression of SREBP family members, SREBP-1 and SREBP-2 SREBP-1 exists in two forms, designated 1a and 1c SREBP-1a is the predominant isoform in most cultured cells,...

Ngày tải lên: 28/03/2014, 22:21

... Determination of the regulation of these genes will require further study of the potentially complex contribution of expression, phosphorylation, and co-factor interaction of wild-type and p53 mutants, and ... Resau J, Bradford C, Carey T, et al.: Genetic and expression profiles of squamous cell carcinoma of the head and neck correlate with cisplatin sensitivity and resistance in cell lines and patients ... in orthologous promoters were 65% for NF-κB, 67% for p53 and AP-1, 73% for STAT3, and 64% for EGR1 These levels of conservation indicated that the majority of predicted TFBSs falling in the orthologous...

Ngày tải lên: 14/08/2014, 07:21

... strains of B subtilis are high secretors and host strains used for successful expression of recombinant proteins are often deleted for genes amyE, aprE, nprE, spoIIAC, srfC and transformed via ... prices (per gram) of recombinant proteins were $375 for human insulin, $23,000 for tPA, $35,000 for human growth hormone, $384,000 for GM-CSF, $450,000 for G-CSF, and $840,000 for EPO All except ... those of the Gram-positive bacilli These are mainly preferred for homologous expression of enzymes such as proteases (for detergents) and amylases (for starch and baking) Some advantages of using...

Ngày tải lên: 08/09/2015, 22:11

... strains of B subtilis are high secretors and host strains used for successful expression of recombinant proteins are often deleted for genes amyE, aprE, nprE, spoIIAC, srfC and transformed via ... prices (per gram) of recombinant proteins were $375 for human insulin, $23,000 for tPA, $35,000 for human growth hormone, $384,000 for GM-CSF, $450,000 for G-CSF, and $840,000 for EPO All except ... rapid expression, ease of culture and high product yields (Swartz, 1996) It is used for massive production of many commercialized proteins This system is excellent for functional expression of non-glycosylated...

Ngày tải lên: 09/09/2015, 12:14

... strains of B subtilis are high secretors and host strains used for successful expression of recombinant proteins are often deleted for genes amyE, aprE, nprE, spoIIAC, srfC and transformed via ... prices (per gram) of recombinant proteins were $375 for human insulin, $23,000 for tPA, $35,000 for human growth hormone, $384,000 for GM-CSF, $450,000 for G-CSF, and $840,000 for EPO All except ... rapid expression, ease of culture and high product yields (Swartz, 1996) It is used for massive production of many commercialized proteins This system is excellent for functional expression of non-glycosylated...

Ngày tải lên: 09/09/2015, 12:34

... produce recombinant therapeutic proteins in plants This study was conducted to help better understand the conditions affecting molecular farming of recombinant proteins The expression of foreign proteins ... can be used to test recombinant protein stability before stable transformation, transient expression methods are unsuitable for the expression of larger amounts of foreign proteins Particle bombardment ... growth factors as a candidate for expression 42 2.5.1 Molecular structure and development of recombinant Angiopoietin (Ang1) 42 2.5.2 Molecular structure and development of recombinant human Vascular...

Ngày tải lên: 12/09/2015, 11:08

... subsequent cloning of the expression cassette of genes of interest The GUS expression cassette was cloned at this HindIII site to yield pASVGUS for its expression in plant cells The expression cassette ... replicate and express foreign genes in cells of monocotyledonous plants Plant Cell 3:247–258 Mor TS, Moon YS, Palmer KE, Mason HS (2003) Geminivirus vectors for high level expression of foreign proteins ... that participate in the control of replication and gene expression The DNA B component encodes proteins required for nuclear trafficking and cellto-cell movement of the viral DNA Relatively few...

Ngày tải lên: 16/09/2015, 17:13

... 41 Cloning and Expression of MBD1 3.5.1 Cloning of MBD1 41 3.5.2 Expression of MBD1 42 CHAPTER RESULTS AND DISCUSSION vi 4.1 Cloning of Fat1 and Atrophin1 43 4.2 Expression of Fat1 and Atrophin1 ... Subcloning Of Fat1 and Atrophin1 3.2.1 Touch up PCR for Fat1 and Atrophin1 33 3.2.2 Double digestion, phenol-chloroform purification and ligation of Fat1 33 3.2.3 Digestion, Phenol Chloroform extraction ... Chloroform extraction and Ligation of Atrophin1 35 3.3 35 PROTEIN Expression And PurificatiON 3.3.1 Expression of Fat1 35 v 3.3.2 Purification of Fat1 36 3.3.2.1 Affinity Purification 36 3.3.2.2...

Ngày tải lên: 05/10/2015, 22:31

... for a much richer understanding of culture than heretofore envisaged by the majority of language teachers 2.2 Common approaches to the teaching of culture In the history of culture teaching different ... textbook among 23 the set of English textbooks for senior high school in Vietnam and the analysis of its cultural content may set an initial step for the analysis of the whole set of high school English ... of frequency and percentage of types of references to Vietnamese and foreign cultures according to Checklist One Types of reference Reference to Vietnamese culture Reference to foreign culture...

Ngày tải lên: 07/09/2013, 12:58