77 resultados para Lipid merging


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A novel method has been developed to easily isolate the mutants with high lipid yield after irradiating oleaginous yeast cells with carbon ions of energy of 80 MeV/u. Pre-selection of the mutants after ion irradiation was performed with culture medium in which the concentration of cerulenin, a potent inhibitor of fatty acid synthetase, was at 8.96 mu mol/l. Afterwards, lipid concentration in the fermentation broth of the pre-selected colonies was estimated by the sulfo-phospho-vanillin reaction instead of the conventional methanol-chloroform extraction. Two mutants with high lipid yield have been successfully selected out by the combined method. This easy and simple method is much less time-consuming but very efficient in the mutant isolation, and it has demonstrated great potential on mutation breeding in oleaginous microorganism.

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In this study, we used a rheological method to study the shape of DNA-cationic lipid complexes and model polyelectrolyte-lipid complexes. We introduced two kinds of anionic polyelectrolytes, sodium polygalacturonate (PGU) and sodium dextran sulfate (DSS), of varying size, as models for DNA. The prepared complexes were incubated under laminar flow conditions. The results show the same quantitative relation between the shape parameter of lipoplexes and the length of anionic polyelectrolytes, including DNA. The rheological behavior of PGU and DSS were similar to that of DNA. (C) 2004 Elsevier Inc. All rights reserved.

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A minicapillary viscometer utilizing <0.5 ml of sample at a volume fraction of <0.1% is described. The calculated a/b of DPPC/DPPG multilamellar liposome was 1.14 as prolate ellipsoids and a/b of dioleoylpropyltrimethyl ammonium methylsulfate-DNA complex at a charge ratio of 4: 1 (+/-) was 3.7 as prolate ellipsoids or 4.9 as oblate ellipsoids. The deviation of shape from perfect sphere is thus expressed quantitatively in more than two significant figures. In these measurement, the necessary amount of DNA is <0.5 mg.

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In this paper, electrochemical surface plasmon resonance (SPR) method was first used to detect enzymatic reaction in bilayer lipid membrane (BLM) based on immobilizing horseradish peroxidase (HRP) in the BLMs supported by the redox polyaniline (PAn) film. By SPR kinetic curve in situ monitoring the redox transformation of PAn film resulted from the reaction between HRP and PAn, the enzymatic reaction of HRP with H2O2, was successfully analyzed by electrochemical SPR spectroscopy.

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Here, we demonstrated dimethyldioctadecylammonium bromide (DODAB), a cationic lipid, bilayer coated Au nanoparticles (AuNPs) could efficiently deliver two types of plasmid DNA into human embryonic kidney cells (HEK 293) in the presence of serum. The transfection efficiency of AuNPs was about five times higher than that of DODAB. The interaction of AuNPs with DNA was characterized with dye intercalation assay and agarose gel electrophoresis. The morphology of the complex of AuNPs with DNA was observed with scanning electron microscope (SEM). The intracellular trafficking of the complex was monitored with transmission electron microscope (TEM).

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In this study, varieties of lipid bilayer-protected gold nanoparticles (AuNPs) were synthesized through a simple wet chemical method, and then the effect of freeze-thawing on the as-prepared AuNPs was investigated. The freeze-thawing process induced fusion or fission of lipid bilayers tethered on the AuNPs. The UV-vis spectra and transmission electron microscopy experiments revealed that the disruption of lipid bilayer structures on the nanoparticles led to the fusion or aggregation of AuNPs.

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A facile phospholipid/room-temperature ionic liquid (RTIL) composite material based on dimyristoylphosphatidylcholine (DMPC) and 1-butyl-3-methylimidazolium hexafluorophosphate ([bmim]PF6) was exploited as a new matrix for immobilizing protein. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were adopted to characterize this composite film. Hemoglobin (Hb) was chosen as a model protein to investigate the composite system. UV-vis absorbance spectra showed that Hb still maintained its heme crevice integrity in this composite film.

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Nonviral vectors are safer than viral systems for gene therapy applications. However, the limited efficacy always prevents their being widely used in clinical practice. Aside from searching new gene nonviral vectors, many researchers focus on finding out new substances to improve the transfection efficiency of existent vectors. In this work, we found a transfection enhancer, nocodazole (NCZ), for dimethyldioctadecylammonium (DODAB, a cationic lipid) bilayer coated gold nanoparticles (AuNPs) mediated gene delivery. It was found that NCZ produces 3-fold transfection enhancement to HEK 293T cells assessed by flow cytometry (FCM). The result was further confirmed by luciferase assay, in which NCZ induced more than 5 times improvement in transfection efficiency after 48 h of transfection. The results from the inductively coupled plasma mass spectrometry (ICP-MS) and FCM showed that NCZ did not affect the internalization of DODAB-AuNPs/DNA complexes. The trafficking of the complexes by transmission electron microscopy (TEM) indicated that the interrupted transportation of the complexes to the lysosomes contributed greatly to the transfection enhancement.