Intravesicle Isothermal DNA Replication

<p>Abstract</p> <p>Background</p> <p>Bacterial and viral DNA replication was previously reconstituted <it>in vitro </it>from component parts <abbrgrp><abbr bid="B1">1</abbr><abbr bid="B2">2</abbr><abbr bid="B3">3</abbr><abbr bid="B4">4</abbr></abbrgrp>. Significant advances in building minimal cell-like structures also have been made recently <abbrgrp><abbr bid="B5">5</abbr><abbr bid="B6">6</abbr><abbr bid="B7">7</abbr></abbrgrp>. Combining the two approaches would further attempts to build a minimal cell-like structure capable of undergoing evolution by combining membrane encapsulation and genome replication. Towards this end, we attempted to use purified genomic replication protein components from thermophilic bacterial sources to copy strands of DNA isothermally within lipid vesicles.</p> <p>Findings</p> <p>Bacterial replication components (such as helicases and DNA polymerases) are compatible with methods for the generation of lipid vesicles. Encapsulation inside phospholipid vesicles does not inhibit the activity of bacterial DNA genome replication machinery. Further the described system is efficient at isothermally amplifying short segments of DNA within phospholipid vesicles.</p> <p>Conclusions</p> <p>Herein we show that bacterial isothermal DNA replication machinery is functional inside of phospholipid vesicles, suggesting that replicating cellular mimics can be built from purified bacterial components.</p

Soft and dispersed interface-rich aqueous systems that promote and guide chemical reactions

Although aqueous solutions are considered to be sustainable, environmentally friendly reaction media, their use is often limited by poor reactant solubility. This limitation can be overcome by converting aqueous solutions into soft, dispersed interface-rich systems such as polyelectrolyte solutions, micellar solutions, oil-in-water microemulsions or vesicle dispersions. All consist of homogeneously distributed dynamic structures that, in a fashion reminiscent of enzymes, provide local environments that are different from the bulk solution. The presence of soft, dispersed interface-rich structures leads to not only selective reaction accelerations but also changes in reaction pathways, whereby chemical reactions are guided towards desired products. Once again, the analogy to enzyme-catalysed transformations is enticing. In this Review, we illustrate the general concepts applied in such systems and illustrate them with selected examples, ranging from enzyme mimics, the preparation of conductive polymers and transition-metal-catalysed organic syntheses on the industrial scale to the chemistry of prebiotic systems.The financial support from the European Cooperation in Science and Technology (COST) action CM1304 for the stimulating meetings on the ‘Emergence and Evolution of Complex Chemical Systems’ is highly appreciated, as well as the financial support from the Swiss National Science Foundation (project No. 200020_150254), the Polish National Science Center through HARMONIA project No. 2014/14/M/ST5/00030 and the Basque Government (project IT 590–13).Peer reviewe