Giant vesicles as microreactors for enzymatic mRNA synthesis.
Giant vesicles have attracted much attention as possible microreactors for the conduction of enzymatic reactions in an artificial, cell-sized compartment. In this context, we demonstrated in the first part of the present work that giant vesicles formed from 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine in an alternating electric field can be made more permeable to Ca(2+) ions or nucleotide triphosphates by addition of ethanol. This methodology is then applied in a second step whereby these giant vesicles are used as microreactors in which mRNA synthesis can occur. The macromolecules (the DNA template and the enzyme T7 RNA polymerase) are microinjected into a selected giant vesicle, while the substrate molecules (nucleotide triphosphates) are added from the external medium. The fact that mRNA synthesis can be detected is a further step towards our aim: the design of a microreactor that can be seen as a model for a protocell.
Fischer
Aline
A
Institute of Polymers, ETH-Zentrum Universitätsstrasse 6, 8092 Zürich, Switzerland.
Franco
Andrea
A
Oberholzer
Thomas
T
D002118:Calcium / Q000378:metabolism*; D017931:DNA Primers / Q000737:chemistry; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000378:metabolism; D000431:Ethanol / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006801:Humans; D008081:Liposomes / Q000737:chemistry*; D008164:Luminescent Proteins / Q000378:metabolism; D008567:Membranes, Artificial; D008845:Microinjections; D010713:Phosphatidylcholines / Q000378:metabolism*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis*
D017931:DNA Primers; D008081:Liposomes; D008164:Luminescent Proteins; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D000431:Ethanol; C034970:bacteriophage T7 induced DNA polymerase; D004259:DNA-Directed DNA Polymerase; D002118:Calcium; C028694:1-palmitoyl-2-oleoylphosphatidylcholine