Phase Separations in Water–Salt Solutions of Polyelectrolyte Complexes Formed by RNA and Polycations: Comparison with DNA Complexes
✍ Scribed by Per-Olof Wahlund; Vladimir A. Izumrudov; Per-Erik Gustavsson; Per-Olof Larsson; Igor Yu. Galaev
- Publisher
- John Wiley and Sons
- Year
- 2003
- Tongue
- English
- Weight
- 131 KB
- Volume
- 3
- Category
- Article
- ISSN
- 1616-5187
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✦ Synopsis
Abstract
Formation of insoluble polyelectrolyte complexes (PECs) between RNA and polycations was followed by measuring the residual RNA absorbance in the solution after separation of the precipitate. The polycations studied were poly(N,N′‐dimethyldiallylammonium) chloride (pendant type) and 2,5‐ionene bromide (integral type) with quaternary amino groups in every monomer unit. The data obtained were compared with the results of analogous studies of DNA‐containing PECs. This study is a part of a project aimed at the specific separation of plasmid DNA from RNA, a major problem in the preparative isolation of plasmid DNA. We thus deliberately chose a heterogenous RNA sample as it represents the RNA present in a real cell extract. In contrast to the exhaustive precipitation of DNA observed at certain φ values, a significant part of RNA was nonprecipitated at any φ = [+]/[−], that is, at any ratio of positively charged quaternary amino groups and negatively charged phosphate groups. The addition of sodium chloride increased the nonprecipitated fraction of RNA. DNA, on the other hand, was completely precipitated by both polycations at φ > 0.7. The less effective precipitation of RNA was probably due to the presence of a considerable fraction of short‐chained molecules, incapable of forming a sufficient cooperative system of salt bonds with the polycation. This assumption was supported by a separate experiment, in which the precipitation behavior of RNA fractions of different molecular masses was investigated. The same tendency, while less pronounced, was also ascertained for PECs formed by polycations with DNA fractions of different molecular masses. The possibility of using the revealed differences between DNA and RNA behavior for effective precipitation procedure useful in bioseparation is discussed. The difference in the precipitation behavior of nucleic acids of different molecular masses means there is a possibility for developing an enzymatic assay for DNAase and RNAase activity.
The relative residual absorbance of RNA (closed symbols) and DNA (open symbols) remaining in solution after precipitation with 2,5‐ionene bromide as a function of the charge ratio, [+]/[−], determined at different concentrations of sodium chloride (M).
magnified imageThe relative residual absorbance of RNA (closed symbols) and DNA (open symbols) remaining in solution after precipitation with 2,5‐ionene bromide as a function of the charge ratio, [+]/[−], determined at different concentrations of sodium chloride (M).