of aav vector plasmids suitable for transient transfection of 293 cells

... virus-infected Vero cells at 12 h p.i Fig 2b shows an increased degree of cytoskeletal thickening (arrow) of similarly infected cells at 16 h p.i It is noted that the formation of cytoskeleton clustering ... mock-infected Vero cells at 36 h p.i (Fig 3a) These control cells also showed a lack of cytoskeletal formation at the periphery of the plasma membrane when compared to infected cells Fig 3b shows ... a distinct lack of filopodia and cytoskeleton formation is obvious The bright spots depict high regions of the cells (b) AFM amplitude scan of West Nile virus – infected Vero cells at 36 h p.i...

Ngày tải lên: 11/08/2014, 00:22

... polymerization of Gag occurs mainly through the formation of an intermediate layer of cup-shaped structures, each made of one hollow hexameric ring of CA domains placed immediately above a stem formed ... evidence for the occurrence of different conformational rearrangements of CA during the assembly of both the immature and the mature capsid of HIV-1 and other retroviruses Induced conformational ... pseudoatomic models of capsid-like particles [30,39]; (b) analyses of the effect of CA mutations on the assembly of mature capsidlike particles (Fig 2E,F) and ⁄ or the formation of viral cores; the...

Ngày tải lên: 18/02/2014, 06:20

... using deletion mutants of the 3¢ terminus of the minus-strand RNA but deletions were made on the basis of the structure of the 5¢UTR of the plus-RNA, the structure of the 3¢-end of the minusstrand ... enzyme) and the Xho1 site of the pET21b vector For construction of an expression vector of the GBV-B NS5B-D19, a 1728 bp DNA fragment was amplified from the sequence coding for the GBV-B-NS5B kindly ... that the 341 nt of the 5¢UTR are needed for efficient replication of HCV RNA in the replicon system Their study showed that the first 125 nt of the 5¢UTR are sufficient for a low level of RNA replication...

Ngày tải lên: 20/02/2014, 01:20

... (GMD) was calculated for each papilloma every week The mean GMDs and the standard error of mean (SEM) for each group was plotted against time for sites infected with 10-2 dilution of infectious CRPV ... Regression of papillomas on the backs of rabbits (NZW) following vaccination with CRPV VLPs Regression of papillomas on the backs of rabbits (NZW) following vaccination with CRPV VLPs A total of nine ... Page of (page number not for citation purposes) Virology Journal 2008, 5:45 http://www.virologyj.com/content/5/1/45 pControl (Fig 1) The regression rate of the papillomas at every site for each...

Ngày tải lên: 20/06/2014, 01:20

... under the control of the SRα promoter in 293Cre cells as described [49] Amplification and rescue of the vector was achieved with the use of the AdLC8cLuc helper virus Both vectors were purified ... throughout the course of the study Assessment of serum ALT after a single dose of adenovirus A dose-dependent, self-limiting hepatotoxicity, often indicated by transient elevation of serum transaminases, ... on each CYP isoform may be the direct result of interaction of capsid proteins with virus receptors and subsequent internalization of virus particles at the cellular level Binding of the fiber...

Ngày tải lên: 20/06/2014, 01:20

... viruses, by transient expression in 293T cells of surface (HA, NA, M2) and internal (Gag, GFP marker genome) viral components Expression of the different viral proteins in producer cells and their ... using dilutions of the viral supernatant that transduced between 0.1% and 5% of GFP-positive cells Due to the use of a FACS method to monitor transduced cells, the determination of GFP-positive ... displayed as the % of neutralisation of the S2 sera compared to the S0 pre-immune sera, i.e., sera harvested before the first inoculation for each mouse, for a 1/100 dilution of these sera Sera...

Ngày tải lên: 20/06/2014, 01:20

... [29] observed that peripheral blood mononuclear cells of calves experimentally infected with F hepatica expressed high amounts of IL-4 but not of IFN-γ mRNA early in the infection indicating ... In F hepatica-resistant rats larvae of F hepatica were coated with antibody and host cells, including eosinophils, neutrophils, macrophages and mast cells, before they were destroyed within the ... F gigantica The mechanism of killing juvenile flukes in F hepatica-resistant rats was identified as the release of high levels of nitric oxide by peritoneal lavage cells [22,23,27] Cattle and...

Ngày tải lên: 07/08/2014, 18:21

... degree of proliferation of synovial lining cells was assessed in at least five points in an area of more than 1.5 cm × 1.5 cm as follows: -, or cells thick; +/-, or cells thick; ++, 5–9 cells ... but the association of virus infection has not yet been studied In the present study we investigated a potential role of HTLV-I infection in the pathophysiology of primary OA For this, we compared ... of synovia of eight HTLV-I carrier patients, and determined the expression of Tax protein by immunohistochemistry Materials and methods cross-sectional study Patients who fulfilled even one of...

Ngày tải lên: 09/08/2014, 01:23

... expression of TNF-α in RAW 264.7 cells Relative TNF-α expression was analyzed by Real-time PCR (C) Suppression of IL-10 expression by various doses of IL-10 shRNA in RAW 264.7 cells (D) RAW 264.7 cells ... irrelevant shRNA for 36 h After 36 h, conditioned medium was collected for analysis of TNF-α expression by ELISA (F) RAW 264.7 cells (2 × 105) were pre-treated with LPS (2 μg/ml) for 90 Cells were ... Journal of Biomedical Science 2010, 17:51 http://www.jbiomedsci.com/content/17/1/51 264.7 cells with pseudoinfection of VLPs-IL-10 shRNA was shown in Fig 2B The cells with the pseudoinfection of VLPs...

Ngày tải lên: 10/08/2014, 05:21

... active for nucleation of assembly, but further addition of dimers requires that some of them be present in an RNA-free conformation [30] Thus the presence of excess operator RNA is inhibitory of ... the structure of the icosahedron Partial reduction of the PP7 VLP results in the appearance of monomers, and multiples of monomers up to the size of hexamers However, VLPs made of single-chain ... Figure of DTT reduction of PP7 and 2PP7 VLPs with varying concentrations SDS polyacrylamide gel electrophoresis of the products of SDS polyacrylamide gel electrophoresis of the products of reduction...

Ngày tải lên: 11/08/2014, 00:22

... temperature of 37°C Subunit fusion increases the stability of coat protein and thereby suppresses these defects, making possible the formation of VLPs Insertion of foreign peptides into one AB-loop of ... and the polylinker of pET3d and the kanamycin resistance determinant of and origin of replication of pET9 Note that detailed information about pET3d and pET9, as well as the plasmids themselves, ... sequences downstream of a Bam HI site near the 3’ end of the coat Page of sequence Replacement of the Kpn I - Bam HI fragment of p2P7K32 with this PCR product resulted in insertion of the Flag peptide...

Ngày tải lên: 11/08/2014, 00:23

... cytokine profile of HPV infection for the induction of IgG2 [28] Detection of the IgG3 subclass was not performed, because rhesus macaques not make IgG3 [18,19] We also measured high levels of serum ... IL-4-producing T-helper cells, high levels of neutralizing Abs and low levels of IFN-γ secreting CD8+ or CD4+ Page of 11 (page number not for citation purposes) Journal of Immune Based Therapies ... the majority of HPV-related clinical disease Recently, we reported a proof -of- concept efficacy study for a prophylactic vaccine composed of HPV 16 L1 virus-like particles (VLP) formulated on...

Ngày tải lên: 11/08/2014, 10:23

... examples of HHV-8 reactivation without evidence of MCD or KS Van der Kuyl et al detected HHV-8 DNA in the PBMCs of 14% of reported cases of asymptomatic HHV-8 infection (median 2, log/ 10E6 cells) ... initiated on chemotherapy for presumed MCD, high levels of HHV-8 DNA were detected A repeat lymph node biopsy finally established the diagnosis of tuberculosis For a description of the methods, please ... nuclear antigen of HHV-8 allows the detection of B cells infected with HHV-8 These cells have undergone plasma cell differentiation and are mainly located in the mantle zone This method of staining...

Ngày tải lên: 11/08/2014, 17:21

... generation of LASV VLP Results LASV gene expression and incorporation in VLP Transient transfection of HEK-293T/17 cells with LASV GPC, NP, and Z gene constructs resulted in high level expression of ... purification of LASV VLP LASV VLP were generated by transfecting HEK-293T/17 cells in 6-well plates (for small scale analysis) or in 15 cm plates (for purification of multi-milligram quantities of VLP) ... micrographs of LASV VLP budding from the surface of HEK-293T/17 cells expressing LASV Z alone or in combination with GPC and NP genes, and high magnification of LASV pseudoparticles Cells expressing...

Ngày tải lên: 12/08/2014, 01:22

... vitro Page of Figure Scheme for the construction of the expression vector p-SMK-Cap with SUMO motif Cloning the Cap gene and construction of a recombinant expression vector A pair of expression ... Only the deletion of the whole NLS domain may allow for the expression of a large amount of the Cap protein However, the NLS plays an important role forming the self assembly of Cap-like particles ... http://www.virologyj.com/content/7/1/166 PCV2-infected pigs Therefore, it is a good candidate antigen for the design of new recombinant vaccines against PCV2 infection and for the development of serologic tests There are several...

Ngày tải lên: 12/08/2014, 04:20

... Thus, the phenotype of our Vif Figure Vif Absence 6of anti-DSB effect of zinc-binding domain mutants of Absence of anti-DSB effect of zinc-binding domain mutants of Vif Sf9 cells were coinfected ... for the difference in DSB response between Sf9 and 5BD.1 cells, as well as other mammalian cells Absence of detectable effect of EnvGp160 or Vpr on the DSB inhibition of VLP assembly in Sf9 cells ... Figure 2of counteracting effect of Vpr on DSB inhibition of HIV-1 VLP assembly and release Absence Absence of counteracting effect of Vpr on DSB inhibition of HIV-1 VLP assembly and release Sf9 cells...

Ngày tải lên: 12/08/2014, 04:21

... after the end of the treatment (0 = no response; = response) cycle of these viruses and their response to therapy [6] Studies of the kinetics of hepatitis C virus (HCV) after initiation of IFN monotherapy ... load decline of log during the first weeks of treatment Patients with SVR had the same viral load decline (3 log) in only four weeks of treatment At the end of week 2, a viral load drop of more than ... reduction of the viral load (in log10) from baseline to week 2, 4, 8, and 12 of treatment, week point time tend to be better than other time points for predicting SVR, with a PPV of 91% and a NPV of...

Ngày tải lên: 13/08/2014, 13:20

... line (HepG2 cells, × 105 cells) was seeded onto each well of a six-well plate until the cells reached 80% confluence (~5 × 105 cells) The cells were infected with × 1010 particles of rAAV-TTR-RB15 ... delivery vectors for prolonged gene expression We sought to use adeno-associated viruses (AAV) for the longevity of transgene expression [4] Furthermore, the AAV vector does not elicit T cells immune ... The expression of self-complementary AAV2-TTR-RB15 (scAAV2-TTR-RB15) vector in HepG2 cells The expression of self-complementary AAV2-TTR-RB15 (scAAV2-TTR-RB15) vector in HepG2 cells (A) The drawings...

Ngày tải lên: 14/08/2014, 19:22

... posttransfection were analyzed for virus release by measuring p25 and vector RNA genome copies and for competence for transduction in 293T cells The efficiency of transfection was similar for ... and 293T cells, using LA34 pseudotyped with VSV-G (LA34/VSV-G) generated in CrFK or 293T cells with p∆env1 as packaging construct The proportions of GFPpositive CrFK and 293T cells days post -transfection ... positive Vector production Vectors were generated in CrFK or 293T cells Briefly, 2.8 × 106 cells were seeded in 10 cm Petri dishes and one day later co-transfected with one of the vector plasmids, ...

Ngày tải lên: 14/08/2014, 19:22