PubMed · 14300212
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R A SCOTT, H A SCHERAGA. 1965-03-15. METHOD FOR CALCULATION INTERNAL ROTATION BARRIERS.. https://doi.org/10.1063/1.1696269
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Polyethylenimine (PEI) is one of the most potent non-viral vectors. We have developed a lactosylated PEI (Lac-PEI) to enhance cell-specific transfection and have shown that Lac-PEI is more efficient than unsubstituted PEI for gene transfer into immortalized cystic fibrosis airway epithelial SigmaCFTE29o-cells. As both intact PEI/plasmid and Lac-PEI/plasmid complexes are found in the cell nucleus, we have investigated the transcription efficiency of the plasmid complexed with PEI or Lac-PEI, according to the polymer nitrogen/DNA phosphate (N/P) ratio (from 0 to 20). The initiation of transgene transcription was analyzed in an acellular nuclease S1 transcription assay. For both PEI and Lac-PEI complexes, transcription efficiency varied with the N/P ratio of the complexes. Transcription inhibition was observed when plasmid DNA was either loosely (N/P<5) or tightly condensed (N/P>15). For an N/P ratio of 5 and up to 15, transcription of the complexed plasmid was as efficient as that of the free plasmid. Similar results were observed when gene expression was studied after nuclear microinjection of the complexes into SigmaCFTE29o-cells. Our study shows that condensation of DNA influences the accessibility of the plasmid to the transcription machinery. Interestingly, the charge ratios that allow the most efficient transcription are those usually known to be the most efficient for gene transfer in vitro and in vivo.
This work deals with the modeling of electrophoretic mobilities of end-charged homopolypeptides. The ionic mobilities of six families of homopolypeptides (polyglycines, poly-L-alanines, poly-L-valines, poly-L-leucines, poly-L-isoleucines, and poly-L-phenylalanines), with polymerization degrees up to 11, have been carefully determined. The electrophoretic frictional coefficients derived from the ionic mobility values were modeled by the hydrodynamic frictional coefficient of an equivalent cylinder. The hydrodynamic modeling allowed the determination of the molecular dimensions of the homopolypeptides in the electrolyte. The results were in good accordance with the expected geometry of the molecules. This approach allows monitoring the change in peptide conformations as a function of the experimental conditions (temperature, nature of the solvent) through the determination of geometrical molecular parameters (total peptide length, lateral radius of the equivalent cylinder, and folding angle). The influence of the bulkiness of the homopolypeptide lateral chain on the conformation is also discussed.
Langmuir and Langmuir-Blodgett (LB) films of a great variety of molecular metal complexes with interesting magnetic or optical properties have been prepared in the last few years. Some of the results obtained by our group and others are summarized in this article.