in vitro mutagenesis of brucella species

... Table 2: In vitro susceptibilities of Brucella melitensis isolates to 11 antibiotics Antibiotics Tetracycline Rifampicin Streptomycin Gentamicin Norfloxacin Ciprofloxacin Levofloxacin Trimethoprim/ ... values of tetracycline, ciprofloxacin, levofloxacin, and amoxicillin/clavulanic acid, interpreted according to the NCCLS criteria for slow growing bacteria, have shown ranges below the breakpoints ... of a kind of antibiotic resistance have been reported [7,8] The aim of this essay was to determine the antibiotic susceptibilities of the Brucella isolates identified in the Laboratory of Clinical...

Ngày tải lên: 08/08/2014, 19:20

... concen- trations of BA, but not of K in combination with auxins, promote shoot proliferation probably by stimulating axillary bud growth Medium to high concentrations of auxin in combination with ... abscission Rooting of P cineraria was obtained only with combinations of K and IBA (0.05 mg/I K, 1-15 mg/I IBA, and mg/I K, 15 mg/I IBA), the greatest number of roots (2.8/ explant) being initiated ... Shoot proliferation of P cineraria was also promoted by high levels of BA, 10 mg/I BA, mg/I NAA resulting in shoots/node K failed to induce multiple shoot production in P cineraria Shoot growth...

Ngày tải lên: 09/08/2014, 04:20

... also protein fragments for mapping of interaction domains The use of in vitro expression approaches to analyze protein products is likely to gain momentum for the analysis of known proteins as well ... functionally analyzed using in vitro assays Furthermore, rapid in vitro protein–protein interaction studies could be performed using this technique where not only full-length version of proteins could be ... original primers Of course in this case the use of an enzyme with 3′-exonuclease proofreading activity should be avoided to protect the differentiating 3′-end An example of the design of original...

Ngày tải lên: 25/10/2013, 22:20

... reaction by using 32P-labeling primer or template in the presence or absence of DNA-binding agents (DDIs and doxorubicin) The DNA bulge binding of both compounds was detected by CD and UV melting experiments ... is formed To gain insight into the binding of drugs to bulge DNA, several bulgecontaining oligonucleotides were selected as binding hosts for DDI-1A and DDI-1B CD spectroscopy of DDI-1A (Fig ... two-base bulge in DNA via stacking of its helically oriented aromatic ring systems on the bulge-flanking base pairs that define the long sides of the triangular prism binding pocket, with its amino sugar...

Ngày tải lên: 07/03/2014, 06:20

... alone did not increase the extent of staining However, in cells that had been cocultured with CDO and a cytotoxic drug, staining was significantly increased, indicating the presence of senescence ... provide a possible model of drug interaction in GEO cells, which involves the complex interaction of proteins involved in cell cycle regulation To recapitulate, combining CDO with cytotoxic chemotherapy ... analysis of BrdU incorporation The degree of incorporation of the thymidine analogue 5-bromo-2¢-deoxyuridine (BrdU; Sigma) in HCT116 and GEO cells was measured by flow cytometry Following culture in...

Ngày tải lên: 07/03/2014, 15:20

... the initial screening is conducted in vitro because of the large number of candidate plant species and the prohibitive cost of screening them in vivo The number of species for in vivo testing ... challenges of extrapolating in vitro findings to in vivo evaluation of plant resources The chapters in this book are written by experts interested in exploring and making better use of plant biodiversity ... remaining larvae into individual labelled petri dishes and stain with a few drops of helminthological iodine 10 Add 50 μL of helminthological iodine to each well on the culture plate 11 Using...

Ngày tải lên: 15/03/2014, 04:20

... (Fig 3E) Interestingly, the inclusion of IKK2 inhibition by SC-514 with our experimentally determined rate constants in the original model resulted in a delay in simulated peak and damping of subsequent ... simulated in silico Combining this method of analysis (in vitro, cell-based and in silico analysis) will facilitate systematic understanding of the underlying properties of this signaling pathway ... = Intensity of nucleus Intensity of cytoplasm Intensity of nucleus Scheme This is a simplication of a cell as seen by analysis software, where the software measures the intensity of NF-jB in...

Ngày tải lên: 16/03/2014, 11:20

... stringent concentration of KCl, there was a correlation between inhibition of endonuclease activity and inhibition of splicing activity indicating that RNA binding can be monitored by measuring ... and its binding affinity is not altered by the presence of DNA in the DNA binding site Interestingly, when nM RNA and 10 nM DNA were added simultaneously to a limiting amount of protein in TK8 buffer ... background In vitro competition assays RNA splicing and DNA cleavage competition experiments both involved a prebinding step in which either DNA or RNA substrates were incubated with I-AniI in a binding...

Ngày tải lên: 17/03/2014, 10:20

... in the GPI-bALP mediated mineralization: effects of Ca2+ concentration To assess the role of Ca2+ in initiating the mineralization process, we modelled the biological process by using 0.8 U of ... presence of anchored GPI-bALP (Fig 1) The presence of HA in the mineral samples was confirmed by In uence of other phosphate substrates in the mineralization process To examine the effects of substrates ... of HA No mineral material was formed in media lacking GPI-bALP Mineral calcium phosphate identification by infrared spectra To identify the quality and the type of mineral formation obtained in...

Ngày tải lên: 23/03/2014, 17:21

... functioning of pSELECT- with an insert, we first deleted the Eco R1 site in the linker by cutting with this enzyme, filling in the ends with Klenow and religating A 792 b.p Hind III fragment containing ... generate many ampicillin resistant colonies starting from single-stranded DNA and following the in vitro fill in reaction outlined in Materials and Methods When the complement of the above oligonucleotide ... by propagating the plasmid in E coli NM522 and infecting with M13K07 helper phage In vitro mutagenesis to remove the Hind Im site was performed by hybridizing an oligonucleotide having the sequence...

Ngày tải lên: 23/03/2014, 22:20

... types (Fig 1A,B) in the presence of 0.2 mM of ubiquinone and, to some extent, of dPQ, duroquinone or vitamin K1 (phylloquinone) In contrast, addition of benzoquinone did not lead to such changes ... by Fisher and Rich [20] for quinone binding sites Although speculative at this stage, this consensus sequence was found in various quinone-binding proteins including AOX [20,21] It should be mentioned ... quinone binding site would be expected to be close to iron binding amino acids, which is the case here ([6], Fig 4) This potential quinone-binding site appears to be conserved in PTOX sequences (including...

Ngày tải lên: 31/03/2014, 07:20

... the background of nonmutated parental DNA, including in vivo incorporation of uracil in the parental strand (3,4), and in vitro incorporation of 5-methyl cytidine or thiophosphates in the synthesized ... also result in nicking at apyrimidinic sites Presumably, this nicking results in preferential selection of the in vitro synthesized strand as template during in vivo replication, which, in turn, ... step of in vitro UDG treatment has been introduced to effectively eliminate the parent strand, and thereby increase the efficacy of From: Methods in Molecular Biology, vol 182: In Vitro Mutagenesis...

Ngày tải lên: 11/04/2014, 06:56

... contains a multiple cloning site flanked by opposmg SP6 and T7 RNA polymerase promoters, inserted into the DNA encoding the 1acZ a-peptide Cloning of a DNA insert into the multiple cloning site ... Preparation of Single-Stranded DNA Template Prepare an overnight culture of cells containing recombinant phagemtd DNA by picking a single antibiotic resistant colony from a fresh plate Inoculate mL of ... still is instrumental to learn about the importance of functional domains or even single amino acids of proteins Of the many useful methods available for site-directed mutagenesis, in this chapter...

Ngày tải lên: 11/04/2014, 09:47

... Effect of molarity and pH on the diffusion profile of ketoprofen .85 Effect of temperature on the diffusion profile of ketoprofen 86 Effect of mass on the diffusion profile of ketoprofen ... variability in percutaneous absorption, the precision of dosing, the reservoir capacity of the skin, heterogeneity and inducibility of the skin in turnover and metabolism, inadequate definition of bioequivalence ... include extrusion of lamellar bodies, loss of nucleus and increasing amount of keratin in the stratum corneum (12) The origins of the cells of the epidermis lie in the basal lamina between the...

Ngày tải lên: 13/04/2014, 09:25

... cells producing TGF-β resulted in the induction of Treg in vivo and in vitro in a murine model [52] Moreover, the blockade of TGF-β reduces Treg induction by the fusion cell vaccine and enhances ... (obtained with following informed consent and approved by our institutional review board obtained) was isolated and maintained in TIL Media I medium with 10% heat-inactivated FCS, mM L-glutamine, ... each vaccination [27,33] Autologous FCs were irradiated, suspended in 0.3 ml normal saline, and underwent up to nine times vaccinations via SC injection in the left inguinal area at 2-week intervals...

Ngày tải lên: 18/06/2014, 15:20

... comparing two different MYXV strains (a wild-type strain (T1) and a cell-cultured attenuated vaccinal strain (SG33) [14]) we verified the stability of the viral tropism in vitro Findings Three bovine ... http://www.virologyj.com/content/4/1/94 information concerning interactions between MYXV and bovine cells is available yet In this study, we characterized the infection of bovine cell lines and bovine peripheral blood ... KOP-R P3 BT MDBK Figure Permissivity of bovine cell lines for myxoma virus Permissivity of bovine cell lines for myxoma virus Bovine cells were maintained in DMEM (KOP-R and BT) or MEM (MDBK)...

Ngày tải lên: 18/06/2014, 18:20

... after adding T cells), fixed in buffered formalin and embedded in paraffin a, b: Staining with H&E c, d: Specific brown staining of apoptotic cells by anti-caspase Ab e, f: In situ end-labeling (ISEL); ... through binding of CCL2 to its corresponding chemokine receptor CCR2 on the T cells This was demonstrated by blocking of T-cell migration in presence of addition of excess chemokine CCL2 in the ... cells resulting in tumor growth inhibition (reviewed in [25] and [1]) In mouse systems, ex vivo transduction of chemokines into tumor cells has provided potent tumor vaccines inducing tumor rejection,...

Ngày tải lên: 18/06/2014, 19:20

... comparing two different MYXV strains (a wild-type strain (T1) and a cell-cultured attenuated vaccinal strain (SG33) [14]) we verified the stability of the viral tropism in vitro Findings Three bovine ... http://www.virologyj.com/content/4/1/94 information concerning interactions between MYXV and bovine cells is available yet In this study, we characterized the infection of bovine cell lines and bovine peripheral blood ... KOP-R P3 BT MDBK Figure Permissivity of bovine cell lines for myxoma virus Permissivity of bovine cell lines for myxoma virus Bovine cells were maintained in DMEM (KOP-R and BT) or MEM (MDBK)...

Ngày tải lên: 20/06/2014, 01:20

... cell lines and/or due to the method used to induce Se deficiency in our in vitro model In studies employing Jurkat E6 cells, the Se-deficient medium comprised of insulin, transferrin and bovine ... re-blotting the same membranes with anti-β-actin to water and oxygen Diminished level of GPx1 results in the accumulation of H2O2, which was assayed using H2DCF-DA, a cell-permeable indicator of intracellular ... http://www.virologyj.com/content/5/1/66 lines in which Se deficiency can be efficiently induced in vitro without compromising cell viability Based on the Se-deficient in vitro and in vivo pathogenesis studies using HIV, H3N2...

Ngày tải lên: 20/06/2014, 01:20

... testing the susceptibility of a wide range of clinical isolates of influenza A viruses to chloroquine, as well as in vivo studies, would be needed to determine the role, if any, of chloroquine in ... pathology of avian influenza infection in humans appears to be mediated by pro-inflammatory cytokines [13] Although we have shown that chloroquine is able to inhibit two reference subtypes of influenza ... chloroquine in the prophylaxis and treatment against influenza infection It would also be interesting to determine the activity of chloroquine against the highly pathogenic H5N1 subtype as this...

Ngày tải lên: 20/06/2014, 01:20