Modification of an amplification reaction in recursively dynamic compartments driven by stirring.
In living systems, biochemical reactions are confined to cellular or subcellular compartments, such as the plasma membrane and the organelles within a cell. These biological compartments are usually subjected to recursive changes, such as combinations of growth, fusion, and division, to constitute repeating cell cycles. In such recursively dynamic compartments, the encapsulated biochemical reaction may exhibit dynamics that differ from those of the static compartment (i.e., test tubes) used in conventional biochemistry experiments. To test this hypothesis in a simplified model, we mechanically stirred femtoliter-sized water-in-oil emulsion droplets so that individual droplets were subjected to repeated coalescence and breakage. We show that recursive dynamics appeared in the emulsion, which were measured by the exponential propagation of a water-soluble dye. The rate of the propagation, μ, was controlled by modulating the pulse-width of stirring in an electromagnetic stirrer. Within this system, we studied the dynamics of an RNA-amplification reaction in recursively increasing reaction compartments at various values of μ. We showed that there was an optimal value of μ that maximized RNA amplification. This effect was explained by the balance between the opposing effects of supply of substrate and the dilution of amplified RNA both resulting from coalescence. Moreover, when we mixed two RNA species with different kinetic properties, we found a preferential amplification for one of the species only in the recursively dynamic emulsion. This effect was partly explained by a separation effect which preferentially amplifies the number of compartments for the molecular specie that can better follow the breakage dynamics of the compartments. The present work demonstrated how the recursive dynamics of compartments modifies the internal biochemical reaction.
ERATO, Japan Science and Technology Agency , 1-5 Yamadaoka, Suita, Osaka 565-0871, Japan.
Tetsuo
T
Ichii
Genya
G
Tanahashi
Hiroaki
H
Suzuki
Tetsuya
T
Yomo
D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009821:Oils / Q000737:chemistry; D013696:Temperature; D014867:Water / Q000737:chemistry
D009821:Oils; D014867:Water