Proto-cell Model Exhibiting Synchronized Self-replication Dynamics of Giant Vesicles and DNA

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1. Self-reproducing giant vesicles composed of anionic membrane molecules A novel membrane molecule with an anionic polar head comprised of hetero-phosphate diester was prepared from 10-(p-formylphenoxy) decanol and dodecanol through an amidite method in a 62 % yield. This anionic membrane molecule constructed nearly uni-lamella giant vesicles in a phosphate buffer solution containing 15% methanol. Moreover the anionic vesicles exhibited self-reproduction dynamics when a membrane-precursor was added (The 88th Annual Meeting of Chemical Society of Japan, 3L3-17).2. Polymerase chain reaction (PCR) in giant vesicles composed of POPC Replication of template DNA consisting of 1229 by that can express GFP was performed inside giant vesicles (GV) of POPC (palmitoyl-oleoyl-phosphatidyl choline) using a PCR technique. After 20 thermal cycles, the fluorescence emitted from SYBR Green I absorbed on the DNA-double strands became clearly visible (two orders increase in average) by fluorescence micr … More oscope. The exact replication of the template DNA was confirmed by PAGE (polyacrylamide gel electrophoresis). Flow-cytometric analysis revealed that ratio of the effective replication of DNA inside GVs was ca 10 % (The 88th Annual Meeting of Chemical Society of Japan, 3C6-25).3. Coupling of dynamics of self-replication of DNA and self-reproduction of giant vesicles For the purpose titled as above, a conjugate molecule consisting of a DNA oligomer (15mer), a spacer, and cholesterol was prepared. The DNA oligomer serves as a primer hybridizing with a template DNA (100mer) inside GV, and cholesterol works as, an anchor to fix the conjugate molecule with the vesicular membrane. The progress of the complimentary replication of the template DNA on the inner surface of vesicular membrane was directly observed by a fluorescent microscope using a primer tagged with a fluorescence-probe, and the replication of the template DNA was confirmed by PAGE analysis. This approach guarantees the possible coupling between the two dynamics, which is crucial for giving rise to an artificial cell model (Soft Matter, 2005). Less