production isolation and purification of viral particles

... 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

... PPV-VP2 FD and PPV-VP2 FD The fragment was about 1840 bp 3.2 Expression and purification of polyhistidine-tagged VP2 of PPV We have previously shown that formation of chimeric VLPs of PPV fusion ... virus particles grown in primary cell cultures of porcine origin The method is both labor intensive and costly, with the additional hazard of requiring the handling of large quantities of infectious ... produce VP2 VLPs of infectious bursal disease virus [19], and parvoviruses including PPV [14] and B19 [15] To further facilitate the use of PPV VP2 for diagnosis and vaccination, the goal of the current...

Ngày tải lên: 11/08/2014, 21:21

... a volume of 30 μl of PBS, as follows: PBS containing μg of CT adjuvant (Sigma), 15 μg of p24, 30 μg of RTB/p24, 15 μg of p24 mixed with μg of CT and, 30 μg of RTB/p24 mixed with μg of CT adjuvant ... weight mass of the predicted translation products from sequences RTB/p24 and p24 was 57 kDa and 34 kDa respectively E coli expression of RTB/p24 and p24 proteins Purification of RTB/p24 and p24 was ... chromatography purification E coli expression of RTB/p24 and p24 HIV proteins Six E coli strains were tested to obtain the best levels of expression and solubility of the RTB/p24 and p24 proteins...

Ngày tải lên: 12/08/2014, 04:21

... polysaccharide One milligram of the partially purified polymer contained 309.7 µg of protein, 160.2 µg of neutral sugar, 58.5 µg of hexosamine, 30.8 µg of uronic acid and 3.8 µg of phosphorus Constituent ... concentration of ppm to estimate the effect of pH After min, A660 was measured and the extent of flocculation was calculated Pronase digestion of extracellular polymer Forty milligrams of the extracellular ... this study were of reagent grade Results and Discussion Extraction and partial purification of extracellular polymer The floc was satisfactorily deflocculated within 10 by sonication of the activated...

Ngày tải lên: 05/09/2013, 09:08

... for cDNA single strand synthesis, 600 ng of RNA, 0.5 lg of oligo(dT) primer and 10 mm of dNTPs were incubated for at 65 °C Subsequently, lL of the first strand buffer X and lL of dithiothreitol ... mixture of other isoforms (B) Gel visualization of proteins eluted Lane 1, molecular weight markers Numbers on the left indicate the Mr values of the standards in thousands Lanes 2–9, lg of peak ... P III ⁄ P IV and a mixture of other isoforms (*), respectively, in the presence (lanes 2–5) or absence (lanes 6–9) of 2-mercaptoethanol (C) Elution profile of P III and P IV Chromatofocusing chromatography...

Ngày tải lên: 18/02/2014, 16:20

... of 50 nmÆmin)1, band-width of 1.0 nm, and a resolution of 0.1 nm in the temperature range between 10 °C and 70 °C, after preincubation for 10 at each temperature Three runs were accumulated and ... of 0.6 lM The data were fitted to Michaelis–Menten curves Squares and dotted line: data and fitting with PHI/PHO ratio of 0.5 Triangles and continuous line: data and fitting with PHI/PHO ratio of ... Asterisks and broken line: data and fitting with PHI/PHO ratio of hypothesis and therefore a direct involvement of PHI in electron transfer This conclusion is consistent with the very low degree of identity...

Ngày tải lên: 21/02/2014, 00:20

... intense band of 61 kDa (NrfA) and a band of weak intensity of 19 kDa (NrfH), confirming its hetero-oligomeric nature (Fig 1, lane 1) However, in the absence of boiling (Fig 1A, lanes and 4) high ... and 4) high molecular mass bands of approximately 110 kDa and > 200 kDa were visible, as well as a faint band at 37 kDa, suggesting the presence of dimers All of the bands stained positively for ... between NrfH and NrfA is discussed The reassessment of previous spectroscopic studies was undertaken, particularly regarding the assignment of spectroscopic and redox potentials of the NrfA hemes...

Ngày tải lên: 21/02/2014, 00:20

... BLAST of peptide sequences of 21 kDa proteins in databases Tryptic peptides were puri®ed by RP-HPLC and some of them were totally or partially analyzed by Edman sequencing (ááá) and/ or by MALDI-TOF ... BAS1 protein of Brassica, spinach, barley and A thaliana, PR1 of Phaseolus and MHF of A thaliana These proteins belong to the 2Cys-Prx subfamily All plant 2Cys-Prx proteins, except BAS1 of barley, ... previously [21] Chloroplastic thioredoxins m and f from spinach were a kind gift of P Schurmann (University È of Neuchatel, Switzerland) Thioredoxins m and h from à Chlamydomonas were puri®ed as...

Ngày tải lên: 08/03/2014, 16:20

... presequence of 19 amino acids The mature protein would have a molecular mass of 41.7 kDa and a pI of 6.34 The YedO protein from E coli and the d-CDes from Arabidopsis showed an overall identity of 36% and ... d-CDes mRNA and protein levels in older plants, during the light phase in a diurnal light ⁄ dark rhythm and under sulfate limitation Results In silico characterization and isolation of the Arabidopsis ... protein has a predicted molecular mass of 43.9 kDa and a pI of 7.2 It contains relatively high amounts of the sulfur amino acids cysteine (four residues) and methionine (10 residues) According...

Ngày tải lên: 16/03/2014, 18:20

... adjustment of linker lengths resulting in various geometric conformations of VH/VL interactions [46], or production of helix-stabilized Fv fragments [47]) and for camelid VHHs (production of a bivalent ... possibility of inter- and intra CDR disulphide cross-links These strategies are summarized in Fig 1B and Table 1, and details of the library construction are given in greater detail in the Materials and ... possibility of disulphide bond formation genetically by replacement of residues Cys22 and Cys82 with alanine and valine, respectively, to give a cysteine minus mutant (12F-11DCys) Use of alanine and...

Ngày tải lên: 17/03/2014, 10:20

... site molecules Y80, Y123 and E177 are shown in green and the position of the two mutated amino acids, F108 and L151, are shown in blue between the aromatic rings of Tyr80 and Tyr123 close to the ... diagram of wild-type RTA structure, and the positions of the altered amino acids of the double mutant, F108 and L151, within the structure are indicated Discussion RTA is the catalytic polypeptide of ... Utilisation of the budding yeast Saccharomyces cerevisiae for the generation and isolation of non-lethal ricin A chain variants Yeast 22, 1287–1297 16 Gemmill TR & Trimble RB (1999) Overview of N- and...

Ngày tải lên: 23/03/2014, 07:20

... Switzerland), thermolysin, chymotrypsin or elastase (Sigma, St Louis, MO, USA)] and Cloning of the bovine MUC15 cDNA by PCR with degenerate primers Isolation of total RNA from the mammary gland of ... PCR screening of bovine and human cDNA libraries showed the presence of MUC15 mRNA in human breast, bovine mammary gland, bovine lymph nodes and lungs of both species (Table 1) Of the identified ... part of the coding sequence was then cloned and sequenced using PCR as described above with 0.4 lM and lM of the specific P7 primer and degenerate primers (P4-P6), respectively The cDNA sequence of...

Ngày tải lên: 24/03/2014, 04:21

... of degenerate and specific primers and by rescreening the cDNA libraries The partial clones of XHIVEP2 and -3 covered 5.9 and 3.2 kb of the respective 3¢ ends and included 3¢-UTRs and poly(A)-tails ... upper strands with [32P]ATP[cP] and T4 polynucleotide kinase The labeled oligomers were annealed by heating to 90 °C an equimolar mixture of the upper and lower strands in reaction buffer and cooling ... -2 and -3) The Xenopus XHIVEPs are characterized with respect to temporal and spatial expression, nuclear import/export activity and DNA binding specificity Materials and methods Isolation and...

Ngày tải lên: 30/03/2014, 13:20

... positive for FSP and negative for cytokeratin 20 in all the cultured and examined cells (Figure 3) Discussion Q-banding analysis of FBs from CD and non-CD subjects demonstrated a normal and stable ... small amount of a soft tissue and have an important bacterial contamination caused by the common intestinal flora, the manual management of the endoscope and endoscopic forceps, and their passage ... majority of affected ones The method of cell culture here described could help in the establishment of novel experiments to study the role of FBs in the pathogenesis of the mucosal damage and to...

Ngày tải lên: 18/06/2014, 15:20

... specific viral mRNA targets AS-ONs inhibit viral protein production by both blocking viral mRNA translation and triggering its degradation Since the discovery of viral inhibition effect of AS-ONs ... targeting on viral mRNA Targeting viral mRNA is one of the most active areas of research and development Several strategies have emerged over the years and are being tested pre-clinically and clinically ... potency and cost Peptide-based drug candidates are limited by insufficient efficacy and unfavorable pharmacokinetics MAbs have increasingly gained favor in large part because of the development of...

Ngày tải lên: 18/06/2014, 22:20

... phylogenies and tracing of evolutionary pathways Furthermore, comparison of multiple representatives of homologous viral proteins can be of importance for understanding the protein structure and function ... review, the utility of CODEHOP-mediated PCR for the identification of novel viruses and the characterization of new viral genes and genomic regions is presented While the focus of this study was ... novel viral genomes Often the linear order of genes within the genomes of related viruses is maintained Thus, the spacing and orientation of specific genes can be predicted in the genomes of Page...

Ngày tải lên: 18/06/2014, 22:20

... Domains of E9 and locations of mutations responsible for drug resistance Domains of E9 and locations of mutations responsible for drug resistance The location of the putative exonuclease and polymerase ... streptomycin and non-essential amino acids Vero Cells were obtained from ATCC and were maintained in MEM with Earl's salts and the addition of 10% FBS and standard concentrations of L-glutamine, ... Mapping and marker rescue of recombinant viruses A Map of PCR fragments in the E9L region that were used for marker rescue mapping experiments and reconstruction of resistant viruses B Results of...

Ngày tải lên: 20/06/2014, 01:20

... interests Luis M Branco, Alex Matschiner, and Darryl B Sampey are co-founders and sole members of the Board of Directors of BioFactura, Inc., and have received salary and other compensation from the company, ... expression system (pMAL Protein Fusion and Purification System Manual, New England BioLabs); and (2) translocation of LASV GP1 and NP to the periplasmic space of E coli was toxic to the host cells ... 0.2 g/L of uridine, g/L of glucose, 0.372 mL/L of Dawes Trace 1, 2.14 mL/L of Dawes Trace 2, 0.072 mL/L of Dawes Trace 3, 0.606 mL/L of M calcium chloride dihydrate (CaCl2-2H2O), 0.30 mL/L of 0.43...

Ngày tải lên: 20/06/2014, 01:20

... the G and P serotype specificities of the YK-1 strain, the sequences of the genes encoding both VP7 and VP4 were determined and compared with those of representatives of established G and P serotypes ... rotavirus RNA segment 11, and (c) schematic diagram of the sequences of gene segment 11 Arrows indicate segment 11 of RRV and YK-1, or rearranged segment 11 of variant 311 The YK-1 and its variant clone ... are known, and the NSP4 of YK-1 can be classified as Group C by comparison of the amino acid sequence (aa 131–148) of the variable portion in the VP4 binding domain of various groups of NSP4 (Figure...

Ngày tải lên: 20/06/2014, 01:20

... specific viral mRNA targets AS-ONs inhibit viral protein production by both blocking viral mRNA translation and triggering its degradation Since the discovery of viral inhibition effect of AS-ONs ... targeting on viral mRNA Targeting viral mRNA is one of the most active areas of research and development Several strategies have emerged over the years and are being tested pre-clinically and clinically ... potency and cost Peptide-based drug candidates are limited by insufficient efficacy and unfavorable pharmacokinetics MAbs have increasingly gained favor in large part because of the development of...

Ngày tải lên: 20/06/2014, 04:20